Hydantoin derivatives useful as kv3 inhibitors

ABSTRACT

The invention provides compounds of formula (I): Said compounds being inhibitors of Kv3 channels and of use in the prophylaxis or treatment of related disorders.

TECHNICAL FIELD

This invention relates to novel compounds, pharmaceutical compositionscontaining them and their use in therapy, in particular in theprophylaxis or treatment of hearing disorders, including hearing lossand tinnitus, as well as schizophrenia, bipolar disorder, epilepsy andsleep disorders.

BACKGROUND TO THE INVENTION

The Kv3 voltage-gated potassium channel family includes four members,Kv3.1, Kv3.2, Kv3.3, and Kv3.4. Genes for each of these subtypes cangenerate multiple isoforms by alternative splicing, producing versionswith different C-terminal domains. Thirteen isoforms have beenidentified in mammals to date, but the currents expressed by thesevariants appear similar (Rudy and McBain, 2001, Trends in Neurosciences24, 517-526). Kv3 channels are activated by depolarisation of the plasmamembrane to voltages more positive than −20 mV; furthermore, thechannels deactivate rapidly upon repolarisation of the membrane. Thesebiophysical properties ensure that the channels open towards the peak ofthe depolarising phase of the neuronal action potential to initiaterepolarisation. Rapid termination of the action potential mediated byKv3 channels allows the neuron to recover more quickly to reachsub-threshold membrane potentials from which further action potentialscan be triggered. As a result, the presence of Kv3 channels in certainneurons contributes to their ability to fire at high frequencies (Rudyand McBain, 2001, Trends in Neurosci. 24, 517-526). Kv3.1-3 subtypes arepredominant in the CNS, whereas Kv3.4 channels are found predominantlyin skeletal muscle and sympathetic neurons (Weiser et al., 1994, J.Neurosci. 14, 949-972). Kv3.1-3 channel subtypes are differentiallyexpressed by sub-classes of interneurons in cortical and hippocampalbrain areas (e.g. Chow et al., 1999, J. Neurosci. 19, 9332-9345; Martinaet al., 1998, J. Neurosci. 18, 8111-8125; McDonald and Mascagni, 2006,Neurosci. 138, 537-547, Chang et al., 2007, J. Comp. Neurol. 502,953-972), in the thalamus (e.g. Kasten et al., 2007, J. Physiol. 584,565-582), cerebellum (e.g. Sacco et al., 2006, Mol. Cell. Neurosci. 33,170-179), and auditory brain stem nuclei (Li et al., 2001, J. Comp.Neurol. 437, 196-218).

Characterisation of mice in which one or more of the Kv3 subtypes hasbeen deleted shows that the absence of Kv3.1 gives rise to increasedlocomotor activity, altered electroencephalographic activity, and afragmented sleep pattern (Joho et al., 1999, J. Neurophysiol. 82,1855-1864). The deletion of Kv3.2 leads to a reduction in seizurethreshold and altered cortical electroencephalographic activity (Lau etal., 2000, J. Neurosci. 20, 9071-9085). Deletion of Kv3.3 is associatedwith mild ataxia and motor deficits (McMahon et al., 2004, Eur. J.Neurosci. 19, 3317-3327). Double deletion of Kv3.1 and Kv3.3 gives riseto a severe phenotype characterised by spontaneous seizures, ataxia, andan increased sensitivity to the effects of ethanol (Espinosa et al.,2001, J. Neurosci. 21, 6657-6665; Espinosa et al., 2008, J. Neurosci.28, 5570-5581).

The known pharmacology of Kv3 channels is limited. Tetraethylammonium(TEA) has been shown to inhibit the channels at low millimolarconcentrations (Rudy and McBain, 2001, Trends in Neurosci. 24, 517-526),and blood-depressing substance (BDS) toxins from the sea anemone,Anemonia sulcata (Diochot et al., 1998, J. Biol. Chem. 273, 6744-6749),have been shown to selectively inhibit Kv3 channels with high affinity(Yeung et al., 2005, J. Neurosci. 25, 8735-8745). In addition tocompounds acting directly on Kv3 channels, agonists of receptors thatactivate protein kinase A (PKA) and protein kinase C (PKC) have beenshown to modulate Kv3-mediated currents in specific brain areas, leadingto a reduction in the ability of the neurons to fire at high frequency(Atzori et al., 2000, Nat. Neurosci. 3, 791-798; Song et al., 2005, NatNeurosci. 8, 1335-1342); these studies suggest that PKA and PKC canspecifically phosphorylate Kv3 channels in a neuron-specific manner,causing a reduction in Kv3-mediated currents.

Bipolar disorder, schizophrenia, anxiety, and epilepsy are seriousdisorders of the central nervous system that have been associated withreduced function of inhibitory interneurons and gamma-amino butyric acid(GABA) transmission (Reynolds et al., 2004, Neurotox. Res. 6, 57-61;Benes et al., 2008, PNAS, 105, 20935-20940; Brambilla et al., 2003, Mol.Psychiatry. 8, 721-37, 715; Aroniadou-Anderjaska et al., 2007, AminoAcids 32, 305-315; Ben-Ari, 2006, Crit. Rev. Neurobiol. 18, 135-144).Parvalbumin positive basket cells that express Kv3 channels in thecortex and hippocampus play a key role in generating feedback inhibitionwithin local circuits (Markram et al., 2004, Nat. Rev. Neurosci. 5,793-807). Given the relative dominance of excitatory synaptic input overinhibitory input to glutamatergic pyramidal neurons in these circuits,fast-firing of interneurons supplying inhibitory input is essential toensure balanced inhibition. Furthermore, accurate timing of inhibitoryinput is necessary to sustain network synchronisation, for example, inthe generation of gamma frequency field potential oscillations that havebeen associated with cognitive function (Fisahn et al., 2005, J. Physiol562, 65-72; Engel et al., 2001, Nat. Rev. Neurosci. 2, 704-716).Notably, a reduction in gamma oscillations has been observed in patientswith schizophrenia (Spencer et al., 2004, PNAS 101, 17288-17293).Consequently, positive modulators of Kv3 channels might be expected toenhance the firing capabilities of specific groups of fast-firingneurons in the brain. These effects may be beneficial in disordersassociated with abnormal activity of these neuronal groups.

In addition, Kv3.2 channels have been shown to be expressed by neuronsof the superchiasmatic nucleus (SCN) the main circadian pacemaker in theCNS (Schulz and Steimer, 2009, CNS Drugs 23 Suppl 2, 3-13).

Hearing loss represents an epidemic that affects approximately 16% ofthe population in Europe and the US (Goldman and Holme, 2010, DrugDiscovery Today 15, 253-255), with a prevalence estimated at 250 millionpeople worldwide (B. Shield, 2006, Evaluation of the social and economiccosts of hearing impairment. A report for Hear-It AISBL:www.hear-it.org/multimedia/Hear_It_Report_October_(—)2006.pdf). As lifeexpectancy continues to increase, so too will the number of peoplesuffering from hearing disorders. Furthermore, it is believed thatmodern lifestyles may exacerbate this burden as the younger generationages. Hearing conditions, including tinnitus have a profound effect onthe quality of life, causing social isolation, depression, work andrelationship difficulties, low self-esteem, and prejudice. Voltage-gatedion channels of the Kv3 family are expressed at high levels in auditorybrainstem nuclei (Li et al., 2001, J. Comp. Neurol. 437, 196-218) wherethey permit the fast firing of neurons that transmit auditoryinformation from the cochlear to higher brain regions. Loss of Kv3.1channel expression in central auditory neurons is observed in hearingimpaired mice (von Hehn et al., 2004, J. Neurosci. 24, 1936-1940), and adecline in Kv3.1 expression may be associated with loss of hearing inaged mice (Jung et al. 2005 Neurol. Res. 27, 436-440). Furthermore,pathological plasticity of auditory brainstem networks is likely tocontribute to symptoms that are experienced by many people sufferingfrom hearing loss of different types. Recent studies have shown thatregulation of Kv3.1 channel function and expression has a major role incontrolling auditory neuron excitability (Kaczmarek et al., 2005,Hearing Res. 206, 133-145), suggesting that this mechanism could accountfor some of the plastic changes that give rise to tinnitus. Morespecifically, a reduction in Kv3-like potassium currents in neurons ofthe dorsal cochlear nucleus has now been observed following acoustictrauma in rats, suggesting that reduced Kv3 function could contribute tothe pathological process that is triggered by damaging noise (Pilati etal., 2011, Hearing Res., doi: 10.1016/j.hearingres.2011.10.008), andsupporting the hypothesis that positive modulation of Kv3 channels inauditory brainstem nuclei could have a therapeutic benefit in patientssuffering from noise-induced hearing loss. Finally, Fragile X syndromeand autism are frequently associated with hypersensitivity to sensoryinput, including auditory stimuli. Recent findings suggest that theprotein coded by the FMR-I gene, whose mutation or absence gives rise toFragile X syndrome, may directly regulate the expression of Kv3.1channels in the auditory brainstem nuclei (Strumbos et al., 2010, J.Neuroscience, in press), suggesting that mis-regulation of Kv3.1channels could give rise to hyperacusis in patients suffering fromFragile X or autism. Consequently, we propose that small moleculemodulators of Kv3 channels in auditory brainstem nuclei could have abenefit in the treatment of disorders of hearing, including tinnitus andauditory hyper-acuity associated with Fragile X syndrome and autism.

SUMMARY OF THE INVENTION

The present invention provides compounds of formula (I):

wherein:

-   -   R₁ is H, or C₁₋₄alkyl, halo, haloC₁₋₄alkyl, CN, C₁₋₄alkoxy,        haloC₁₋₄alkoxy;    -   R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or        halo;    -   R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo; or R₃ is absent;    -   R₁₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo; or R₁₃ is absent;    -   A is a 5 or 6 membered saturated or unsaturated heterocycle,        with at least one O atom; which heterocycle is optionally fused        with a cyclopropyl group to form a tricycle when considered        together with the phenyl;    -   X is C or N;    -   Y is C or N;    -   R₄ is C₁₋₄ alkyl;    -   R₅ is H, Deuterium, C₁₋₄ alkyl;    -   or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly;    -   wherein R₂ and R₃ may be attached to the same or a different        ring atom; and wherein R₂ may be attached to a fused ring atom.

Compounds of formula (I) may be provided in the form of apharmaceutically acceptable salt and/or solvate. In one embodiment ofthe invention a compound of formula (I) is provided in the form of apharmaceutically acceptable salt.

The compounds of formula (I) may be used as medicaments, in particularfor the prophylaxis or treatment of hearing disorders, including hearingloss and tinnitus, as well as schizophrenia, bipolar disorder, epilepsyand sleep disorders.

Further, there is provided a method for the prophylaxis or treatment ofhearing disorders, including hearing loss and tinnitus, as well asschizophrenia, bipolar disorder, epilepsy and sleep disorders byadministering to a subject a compound of formula (I).

Compounds of formula (I) may be used in the manufacture of a medicamentfor the prophylaxis or treatment of hearing disorders, including hearingloss and tinnitus, as well as schizophrenia, bipolar disorder, epilepsyand sleep disorders.

Also provided are pharmaceutical compositions containing a compound offormula (I) and a pharmaceutically acceptable carrier or excipient.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compounds of formula (I):

wherein:

-   -   R₁ is H, or C₁₋₄alkyl, halo, haloC₁₋₄alkyl, CN, C₁₋₄alkoxy,        haloC₁₋₄alkoxy;    -   R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or        halo;    -   R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo;    -   R₁₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo;    -   A is a 5 or 6 membered saturated or unsaturated heterocycle,        with at least one O atom; which heterocycle is optionally fused        with a cyclopropyl group to form a tricycle when considered        together with the phenyl;    -   X is C or N;    -   Y is C or N;    -   R₄ is C₁₋₄ alkyl;    -   R₅ is H, Deuterium, C₁₋₄ alkyl;    -   or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly;    -   wherein R₂ and R₃ may be attached to the same or a different        ring atom; and wherein R₂ may be attached to a fused ring atom.

Suitably R₁ is H or methyl. In one embodiment of the invention R₁ is H.In a second embodiment the invention R₁ is C₁₋₄alkyl, in particular R₁is methyl.

Suitably R₂ is H, F, methyl, ethyl, isopropyl or a C₃ spiro group. Inone embodiment of the invention R₂ is H. In a second embodiment of theinvention R₂ is C₁₋₄alkyl, in a particular example of this embodiment R₂is methyl, in a further example of this embodiment R₂ is ethyl and inanother example of this embodiment R₂ is propyl (e.g. isopropyl). In athird embodiment of the invention R₂ is a C₃ spiro group. In a fourthembodiment of the invention R₂ is a C₄ spiro group. In a fifthembodiment of the invention R₂ is halo, in particular fluoro.

Suitably R₃ is H, F, methyl or ethyl. In one embodiment of the inventionR₃ is H. In a second embodiment of the invention R₃ is C₁₋₄ alkyl, in aparticular example of this embodiment R₃ is methyl, in a further exampleof this embodiment R₃ is ethyl. In a third embodiment of the inventionR₃ is halo, in particular fluoro. The skilled person will appreciatethat, depending on the size, presence of heteroatoms and the degree ofunsaturation of the A ring, R₃ may be absent. Consequently, in anotherembodiment of the invention R₃ is absent.

Suitably, R₃ may be H, F, methyl or ethyl and R₂ may be H, F, methyl,ethyl, isopropyl or C₃₋₄ spiro carbocycly. In particular, R₃ may be H,F, methyl or ethyl and R₂ may be H, F, methyl, ethyl, isopropyl or C₃spiro carbocycly. In certain embodiments R₃ is H and R₂ is H, methyl,ethyl, isopropyl or C₃₋₄ spiro carbocycly. In other embodiments, R₃ ismethyl or ethyl and R₂ is methyl or ethyl, in one example of thisembodiment R₃ and R₂ are both methyl (such as attached to the same ringcarbon atom), in a second example of this embodiment R₃ and R₂ are bothethyl (such as attached to the same ring carbon atom). In furtherembodiments R₃ and R₂ are both fluoro (such as attached to the same ringcarbon atom).

Suitably R₁₃ is may be H, F or methyl. In one embodiment of theinvention R₁₃ is H. In a second embodiment of the invention R₁₃ is C₁₋₄alkyl, in a particular example of this embodiment R₁₃ is methyl. In athird embodiment of the invention R₁₃ is halo, in particular fluoro. Theskilled person will appreciate that, depending on the size, presence ofheteroatoms and the degree of unsaturation of the A ring, R₁₃ may beabsent. Consequently, in another embodiment of the invention R₁₃ isabsent.

In one embodiment of the invention A is a 5 membered saturated orunsaturated heterocycle, with at least one O atom; which heterocycle isoptionally fused with a cyclopropyl group to form a tricycle whenconsidered together with the phenyl. In a second embodiment of theinvention A is a 6 membered saturated or unsaturated heterocycle, withat least one O atom; which heterocycle is optionally fused with acyclopropyl group to form a tricycle when considered together with thephenyl.

In certain embodiments the ring A contains one heteroatom. In otherembodiments the ring A contains two heteroatoms (e.g. two oxygen atoms,alternatively one oxygen atom and one nitrogen atom).

Suitably, A is dihydrofuran, isoxazole, dihydropyran, 1,3-dioxolane,1,3-oxazine or dihydropyran fused with a cyclopropyl group. In oneembodiment of the invention A is dihydrofuran. In a second embodiment ofthe invention A is dihydropyran. In a third embodiment the invention Ais dihydrofuran fused with a cyclopropyl group. In a fourth embodimentthe invention A is dihydropyran fused with a cyclopropyl group. In afifth embodiment of the invention A is dihydrofuran, isoxazole ordihydropyran. In a sixth embodiment of the invention A is dihydrofuran,isoxazole or dihydropyran, fused with a cyclopropyl group. In a seventhembodiment of the invention A is 1,3-oxazine. In an eighth embodiment ofthe invention A is 1,3-dioxolane.

When A contains a 5 membered heterocyle containing one heteroatom,suitably the oxygen atom is located in the meta position relative to thephenyl ring.

When A contains a 5 membered heterocyle containing one heteroatom,suitably the heterocyle is dihydrofuran.

When A contains a 6 membered heterocyle containing one heteroatom,suitably the oxygen atom is located in the meta position relative to thephenyl ring.

When A contains a 6 membered heterocyle containing one heteroatom,suitably the heterocyle is dihydropyran.

When A is a five membered ring, in particular embodiments of theinvention one of R₂, R₃ and R₁₃ is H and the others are both methyl, forexample one of R₂, R₃ and R₁₃ is H and the others are both methylattached to the same ring carbon. Alternatively when A is a fivemembered ring, R₂ is a C₃ spiro group and R₃ and R₁₃ are both H.

When A is a five membered ring fused with a cyclopropyl group, suitablyR₂, R₃ and R₁₃ are all H.

When A is a six membered ring, in particular embodiments of theinvention one of R₂, R₃ and R₁₃ is methyl and the others are both H.Alternatively when A is a six membered ring, R₂ is a C₃ spiro group andR₃ and R₁₃ are both H.

When A is a five membered ring fused with a cyclopropyl group, suitablyR₂, R₃ and R₁₃ are all H.

When A is a six membered ring fused with a cyclopropyl group, suitablyR₂, R₃ and R₁₃ are all H.

In one embodiment of the invention X is C and Y is C. In a secondembodiment of the invention X is N and Y is C. In a third embodiment ofthe invention X is N and Y is N.

Suitably, R₄ is methyl, ethyl, isopropyl or t-butyl. In one embodimentof the invention R₄ is methyl. In another embodiment of the invention R₄is ethyl. In a further embodiment of the invention R₄ is propyl, such isisopropyl. In a yet further embodiment of the invention R₄ is butyl,such as t-butyl.

Suitably, R₅ is H or methyl. In one embodiment of the invention R₅ is H.In a second embodiment of the invention R₅ is C₁₋₄alkyl, in particularR₅ is methyl.

In one embodiment of the invention R₄ and R₅ together form a C₃ spirocarbocycle. In a second embodiment of the invention R₄ and R₅ togetherform a C₄ spiro carbocycle. In a further embodiment of the invention R₄is methyl and R₅ is methyl. In an embodiment of particular interest, R₄is ethyl and R₅ is methyl.

Suitably, R₄ and R₅ have the stereochemical arrangement:

In one embodiment of the invention, R₅ is H and the R₄ substituent is inthe S configuration.

In one embodiment of the invention R₄ is methyl, R₅ is methyl, X is Nand Y is C, such as where A is dihydrofuran, in particular where A isdihydrofuran and R₁ is H, especially where A is dihydrofuran, R₁ is Hand R₂ is a C₃ spiro group.

Compounds of formula (I), or any subset thereof including compounds offormula (Ib) and compounds of formula (Ic) may optionally be provided inthe form of a pharmaceutically acceptable salt and/or solvate. In oneembodiment of the invention a compound of formula (I) is provided in theform of a pharmaceutically acceptable salt. In a second embodiment ofthe invention a compound of formula (I) is provided in the form of apharmaceutically acceptable solvate. In a third embodiment of theinvention a compound of formula (I) is not in the form of a salt orsolvate.

In a further aspect, the invention provides a compound of formula (Ib);

wherein:

-   -   R₁ is H, or C₁₋₄alkyl, halo, haloC₁₋₄alkyl, CN, C₁₋₄alkoxy,        haloC₁₋₄alkoxy;    -   R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or        halo;    -   R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo;    -   A is a 5 or 6 membered saturated or unsaturated heterocycle,        with at least one O atom; which heterocycle is optionally fused        with a cyclopropyl group to form a tricycle when considered        together with the phenyl;    -   X is C or N;    -   Y is C or N;    -   R₄ is C₁₋₄ alkyl;    -   R₅ is H, Deuterium, C₁₋₄ alkyl;    -   or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly;    -   wherein R₂ and R₃ may be attached to the same or a different        ring atom; and wherein R₂ may be attached to a fused ring atom;    -   or a pharmaceutically acceptable salt thereof.

Also provided is a compound of formula (Ic)

wherein:

-   -   R₁ is H, or C₁₋₄alkyl;    -   R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or        halo;    -   R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo;    -   A is a 5 or 6 membered saturated or unsaturated heterocycle,        with at least one O atom; which heterocycle is optionally fused        with a cyclopropyl group to form a tricycle when considered        together with the phenyl;    -   X is C or N;    -   Y is C or N;    -   R₄ is C₁₋₄ alkyl;    -   R₅ is H, Deuterium, C₁₋₄ alkyl;    -   or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly;    -   wherein R₂ and R₃ may be attached to the same or a different        ring atom;    -   and wherein R₂ may be attached to a fused ring atom;    -   or a pharmaceutically acceptable salt thereof.        In respect of the compounds of formula (Ib) and (Ic):

In one embodiment of the invention R₁ is H.

In one embodiment of the invention R₁ is C₁₋₄alkyl. In anotherembodiment of the invention R₁ is methyl.

In one embodiment of the invention R₂ is H.

In one embodiment of the invention R₂ is C₁₋₄alkyl. In anotherembodiment R₂ is methyl. In a further embodiment R₂ is ethyl. In a yetfurther embodiment R₂ is propyl.

In one embodiment of the invention R₂ is a C₃ spiro group.

In one embodiment of the invention R₃ is H.

In one embodiment of the invention R₃ is C₁₋₄ alkyl. In anotherembodiment of the invention R₃ is methyl.

In one embodiment of the invention A is tetrahydrofuran, isoxazole ortetrahydropyran.

In one embodiment of the invention A is tetrahydrofuran, isoxazole ortetrahydropyran, fused with a cyclopropyl group.

In one embodiment of the invention X is C and Y is C.

In one embodiment of the invention X is N and Y is C.

In one embodiment of the invention X is N and Y is N.

In one embodiment of the invention R₄ is methyl. In another embodimentof the invention R₄ is ethyl. In a further embodiment of the inventionR₄ is propyl. In a yet further embodiment of the invention R₄ is butyl.

In one embodiment of the invention R₅ is H.

In one embodiment of the invention R₅ is C₁₋₄alkyl. In anotherembodiment of the invention R₅ is methyl.

In one embodiment of the invention R₄ and R₅ together form a C₃ spirocarbocycle.

In one embodiment of the invention R₄ and R₅ together form a C₄ spirocarbocycle.

In one embodiment of the invention, R₅ is H and the R₄ subsituant is inthe S configuration.

In one embodiment of the invention R₄ is methyl and R₅ is methyl.

In one embodiment of the invention R₄ is methyl, R₅ is methyl, X is Nand Y is C.

In one embodiment of the invention R₄ is methyl, R₅ is methyl, X is N, Yis C and A is tetrahydrofuran.

In one embodiment of the invention R₄ is methyl, R₅ is methyl, X is N, Yis C, A is tetrahydrofuran and R₁ is H.

In one embodiment of the invention R₄ is methyl, R₅ is methyl, X is N, Yis C, A is tetrahydrofuran, R₁ is H and R₂ is a C₃ spiro group.

In one embodiment of the invention the compound is selected from thegroup consisting of:

-   (5R)-3-[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]-5-methyl-2,4-imidazolidinedione;-   (5R)-5-methyl-3-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-2,4-imidazolidinedione;-   (5R)-3-{4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-{6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)-2,4-imidazolidinedione;-   (5R)-3-{4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-2,4-imidazolidinedione;-   (5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-ethyl-2,4-imidazolidinedione;-   7-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5,7-diazaspiro[3.4]octane-6,8-dione;-   6-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-4,6-diazaspiro[2.4]heptane-5,7-dione;-   3-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   (5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-(1,1-dimethylethyl)-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione;-   5,5-dimethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione;-   (5R)-5-ethyl-5-methyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-(6-{[(3S/R)-3-methyl-1,3-dihydro-2-benzofuran-4-yl]oxy}-3-pyridinyl)-2,4-imidazolidinedione    (diastereoisomeric mixture);-   (5R)-5-ethyl-3-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (diastereoisomers 1 and 2);-   (5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (diastereoisomeric mixture);-   (5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (diastereoisomers 1 and 2);-   5,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (racemate mixture);-   5,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (enantiomers 1 and enantiomer 2);-   5,5-dimethyl-3-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione;-   5,5-dimethyl-3-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (enantiomer 1 and enantiomer 2);-   (5R)-5-ethyl-5-methyl-3-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-2,4-imidazolidinedione;-   3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5,5-dimethyl-2,4-imidazolidinedione;-   (5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-(1-methylethyl)-2,4-imidazolidinedione;-   (5R)-3-{6-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-2,4-imidazolidinedione;-   (5R)-3-[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]-5-ethyl-5-methyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-{6-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (racemate mixture, enantiomer 1, enantiomer 2);-   (5R)-5-ethyl-5-methyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione    (diastereoisomeric mixture, diastereoisomer 1, diastereoisomer 2);-   (5R)-5-ethyl-5-methyl-3-[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridinyl]-2,4-imidazolidinedione    (diastereoisomeric mixture, diastereoisomer 1, diastereoisomer 2);-   3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione    (racemate mixture, enantiomer 1, enantiomer 2);

or a pharmaceutically acceptable salt thereof.

In another embodiment of the invention the compound is selected from thegroup consisting of:

-   (5R)-5-ethyl-5-methyl-3-[2-(4-methylchroman-5-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione    (diastereoisomeric mixture, diastereoisomer 1, diastereoisomer 2);-   (5R)-5-ethyl-5-methyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione;-   (5R)-3-[2-(3,3-dimethylisochroman-5-yl)oxypyrimidin-5-yl]-5-ethyl-5-methyl-imidazolidine-2,4-dione;-   (5R)-5-ethyl-5-methyl-3-[2-(7-methylspiro[1H-isobenzofuran-3,1′-cyclobutane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione;-   (5R)-5-ethyl-5-methyl-3-{2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-2,4-imidazolidinedione;-   (5R)-3-{2-[(2,2-difluoro-7-methyl-1,3-benzodioxol-4-yl)oxy]-5-pyrimidinyl}-5-ethyl-5-methyl-2,4-imidazolidinedione;-   (5R)-3-{2-[(2,2-difluoro-1,3-benzodioxol-4-yl)oxy]-5-pyrimidinyl}-5-ethyl-5-methyl-2,4-imidazolidinedione;-   (5R)-5-ethyl-5-methyl-3-{2-[(2,4,4-trimethyl-4H-3,1-benzoxazin-5-yl)oxy]-5-pyrimidinyl}-2,4-imidazolidinedione;-   5,5-dimethyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione;-   3-[2-(3,3-dimethylisochroman-5-yl)oxypyrimidin-5-yl]-5,5-dimethyl-imidazolidine-2,4-dione;-   5,5-dimethyl-3-[2-(7-methylspiro[1H-isobenzofuran-3,1′-cyclobutane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione;-   (5R)-5-ethyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione;-   (5R)-5-ethyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione;-   (5R)-5-ethyl-3-{6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione;-   (5R)-5-ethyl-3-{2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-2,4-imidazolidinedione;-   (5R)-5-ethyl-5-methyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione;-   (5R)-3-[6-(3,3-dimethylisochroman-5-yl)oxy-3-pyridyl]-5-ethyl-5-methyl-imidazolidine-2,4-dione;-   (5R)-3-[6-[(3,3-diethyl-1H-isobenzofuran-4-yl)oxy]-3-pyridyl]-5-ethyl-5-methyl-imidazolidine-2,4-dione;-   (5R)-5-ethyl-5-methyl-3-[6-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]-3-pyridyl]imidazolidine-2,4-dione;-   (5R)-3-{6-[(3,3-dimethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-5-methyl-2,4-imidazolidinedione;-   5,5-dimethyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione;

or a pharmaceutically acceptable salt thereof.

For the avoidance of doubt, the embodiments of any one feature of thecompounds of the invention may be combined with any embodiment ofanother feature of compounds of the invention to create a furtherembodiment.

The term ‘halo’ or ‘halogen’ as used herein, refers to a fluorine,chlorine, bromine or iodine atom. Particular examples of halo arefluorine and chlorine, especially fluorine.

When the compound contains a C₁₋₄alkyl group, whether alone or formingpart of a larger group, e.g. C₁₋₄alkoxy, the alkyl group may be straightchain, branched, cyclic, or a combination thereof. Examples of C₁₋₄alkylare methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, cyclopropyl and cyclobutyl. A particular group of exemplaryC₁₋₄alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl, tert-butyl. An example of C₁₋₄alkoxy is methoxy.

The term ‘haloC₁₋₄alkyl’ as used herein, includes straight chain,branched chain or cyclic alkyl groups containing 1 to 4 carbon atomssubstituted by one or more halo atoms, for example fluoromethyl,difluoromethyl and trifluoromethyl. A particular group of exemplaryhaloC₁₋₄ alkyl include methyl and ethyl groups substituted with one tothree halo atoms, in particular one to three fluoro atoms.

The term ‘haloC₁₋₄alkoxy’ as used herein, includes straight chain,branched chain or cyclic alkoxy groups containing 1 to 4 carbon atomssubstituted by one or more halo atoms, for example fluoromethoxy,difluoromethoxy and trifluoromethoxy. A particular group of exemplaryhaloC₁₋₄ alkyl include methoxy and ethoxy groups substituted with one tothree halo atoms, in particular one to three fluoro atoms.

The term ‘5 or 6 membered saturated or unsaturated heterocycle, with atleast one O atom’ includes for example furan, oxazole, isoxazole,oxadiazole, tetrahydrofuran, pyran, tetrahydropyran, dioxolane, dioxan,morpholine, and oxazoline.

It will be appreciated that for use in medicine the salts of thecompounds of formula (I) should be pharmaceutically acceptable. Suitablepharmaceutically acceptable salts will be apparent to those skilled inthe art. Pharmaceutically acceptable salts include those described byBerge, Bighley and Monkhouse J. Pharm. Sci. (1977) 66, pp 1-19. Suchpharmaceutically acceptable salts include acid addition salts formedwith inorganic acids e.g. hydrochloric, hydrobromic, sulphuric, nitricor phosphoric acid and organic acids e.g. succinic, maleic, acetic,fumaric, citric, tartaric, benzoic, p-toluenesulfonic, methanesulfonicor naphthalenesulfonic acid. Other salts e.g. oxalates or formates, maybe used, for example in the isolation of compounds of formula (I) andare included within the scope of this invention.

Certain of the compounds of formula (I) may form acid addition saltswith one or more equivalents of the acid. The present invention includeswithin its scope all possible stoichiometric and non-stoichiometricforms.

The compounds of formula (I) may be prepared in crystalline ornon-crystalline form and, if crystalline, may optionally be solvated,e.g. as the hydrate. This invention includes within its scopestoichiometric solvates (e.g. hydrates) as well as compounds containingvariable amounts of solvent (e.g. water).

It will be understood that the invention includes pharmaceuticallyacceptable derivatives of compounds of formula (I) and that these areincluded within the scope of the invention.

As used herein “pharmaceutically acceptable derivative” includes anypharmaceutically acceptable ester or salt of such ester of a compound offormula (I) which, upon administration to the recipient is capable ofproviding (directly or indirectly) a compound of formula (I) or anactive metabolite or residue thereof.

It is to be understood that the present invention encompasses allisomers of formula (I) and their pharmaceutically acceptablederivatives, including all geometric, tautomeric and optical forms, andmixtures thereof (e.g. racemic mixtures). Where additional chiralcentres are present in compounds of formula (I), the present inventionincludes within its scope all possible diastereoisomers, includingmixtures thereof. The different isomeric forms may be separated orresolved one from the other by conventional methods, or any given isomermay be obtained by conventional synthetic methods or by stereospecificor asymmetric syntheses.

The subject invention also includes isotopically-labeled compounds whichare identical to those recited in formula (I) but for the fact that oneor more atoms are replaced by an atom having an atomic mass or massnumber different from the atomic mass or mass number most commonly foundin nature. Examples of isotopes that can be incorporated into compoundsof the invention include isotopes of hydrogen, carbon, nitrogen, oxygen,fluorine, iodine and chlorine such as ³H, ¹¹C, ¹⁴C, ¹⁸F, ¹²³I or ¹²⁵I.Another isotope of interest is ¹³C.

Compounds of the present invention and pharmaceutically acceptable saltsof said compounds that contain the aforementioned isotopes and/or otherisotopes of other atoms are within the scope of the present invention.Isotopically labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H or ¹⁴C have beenincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i.e. ³H, and carbon-14, i.e. ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.¹¹C and ¹⁸F isotopes are particularly useful in PET (positron emissiontomography).

Since the compounds of formula (I) are intended for use inpharmaceutical compositions it will readily be understood that they areeach preferably provided in substantially pure form, for example atleast 60% pure, more suitably at least 75% pure and preferably at least85%, especially at least 98% pure (% are on a weight for weight basis).Impure preparations of the compounds may be used for preparing the morepure forms used in the pharmaceutical compositions.

According to a further aspect of the present invention there is provideda process for the preparation of compounds of formula (I) andderivatives thereof. The following schemes detail some synthetic routesto compounds of the invention. In the following schemes reactive groupscan be protected with protecting groups and deprotected according towell established techniques.

In general, the compounds of formula (I) may be made according to theorganic synthesis techniques known to those skilled in this field, aswell as by the representative methods set forth below, those in theExamples and modifications thereof.

Compounds of formula (I), and salts and solvates thereof, may beprepared by the general methods outlined hereinafter. In the followingdescription, the groups A, R₁, R₂, X, Y, R₃, R₄ and R₅ have the meaningsas previously defined for compounds of formula (I) unless otherwisestated.

step (ii): Compounds of formula (I) can be prepared by cyclization ofcompounds of formula (II) in a solvent e.g. dichloromethane with acarbonylating agent e.g. triphosgene preferentially prediluted in thesame solvent and added in a second time at 0° C. in presence of a basee.g. triethylamine. In some cases, ethyl acetate could be used as asolvent. Optionally a catalytic amount of DMAP can be added.

step (i): Compounds of formula (II) can be prepared from compounds offormula (III) by removal of the BOC protective group in acidicconditions e.g. TFA in a solvent e.g. dichloromethane at approximately0° C. or RT.

Compounds of formula (I), wherein X═Y═N or (X═C, Y═N) or (X═N, Y═C) andR₄ and R₅ are not H, can be prepared by nucleophilic aromaticsubstitution. In this reaction a halo-pyridyl or halo-pyrimidylderivative of formula (VII) wherein typically Z=Cl and a phenol offormula (IX) are reacted in the presence of a base such as potassiumcarbonate in a suitable solvent, e.g. in N,N-dimethylformamide or inacetonitrile, with conventional heating or microwave heating.

step (iii): Compounds of formula (I) wherein R₄ and R₅ are not H can beprepared by reaction of a urea of the type produced by step (ii) asshown above and a base such as sodium methoxide in a solvent such asMethanol at temperature ranging from 0° C. to 60° C.

step (ii): The urea product of step (ii) as shown above can be preparedby reaction of anilines of formula (IV) and amino esters (hydrochloridesalt) of formula (XII) in a suitable solvent, e.g. dichloromethane orethyl acetate, with a carbonylating agent, e.g. triphosgene,preferentially prediluted in the same solvent in presence of a base,e.g. triethylamine or diisopropylethylamine, at temperature ranging from0° C. to 60° C., optionally adding a catalytic or stechiometric amountof DMAP.

step (i): Amino esters (hydrochloride salt) of formula (XII) (if notcommercially available) can be prepared from commercially availableamino acids (hydrochloride salt) of formula (XIII) by reaction withmethanol in presence of a catalytic or stechiometric amount of thyonylchloride at temperature ranging from r.t. to reflux.

step (i): Compounds of formula (II) can be prepared from anilines offormula (IV) and amino acids (as free base or hydrochloride salt) offormula (VI) by amidic coupling in the presence of a coupling agent e.g.T3P in a solvent such as ethyl acetate, acetonitrile or a mixture ofthem.

step (ii): Compounds of formula (III) can be prepared from anilines offormula (IV) and N-protected amino acids of formula (V) by amidiccoupling in the presence of a base e.g. DIPEA and of a coupling agente.g. HATU, TBTU in a solvent such as N,N-dimethylformamide.

step (i): Some N-Boc protected amino acids of formula (V) arecommercially available e.g.N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine from for exampleAldrich, N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine from for exampleAldrich, (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acidfrom for example Bachem UK Ltd,N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline from for example Nagase& Co Ltd.

N-protected amino acids of formula (V) can also be prepared fromcompounds of formula (VI) for example with Boc-anhydride in presence ofa base e.g. aqueous NaHCO₃, aqueous sodium hydroxide in a solvent suchas THF, methanol, dioxane. Many descriptions are available in theliterature (for example Tetrahedron, 2006, 62(42), 9966-9972)

Anilines of formula (IV) can be prepared from the nitro compounds (VII).Suitable reactions conditions to transform (VII) into (IV) are forexample:

-   -   reduction in presence of Fe powder and ammonium chloride in a        solvent such as a mixture THF/water for example at room        temperature    -   reduction with tin chloride hydrate in a solvent such as ethanol        with heating for example at reflux.

Anilines of formula (IVa), wherein R₂ is H, C₁₋₄ alkyl, C₃-C₄ spirocarbocycly and R₃ is H, C₁₋₄ alkyl and (X,Y) is not (N,N) can beprepared from the nitro compounds (VIIa) with the conditions describedon Scheme 3 or also with the following conditions:

-   -   reduction with hydrazine hydrate and a catalytic amount of Pd/C        in a solvent such as ethanol with heating for example at reflux.

Compounds of formula (VIIb) wherein X═Y═C or (X═C, Y═N) or (X═N, Y═C)can be prepared by nucleophilic aromatic substitution. In this reactionare used a nitro derivative of formula (VIIIb) wherein Z=F (usually when[X═C, Y═C]) or Z=Cl (usually when [X═N, Y═C] or [X═C, Y═N]) and a phenolof formula (IX) in presence of a base such as potassium carbonate in asolvent e.g. in N,N-dimethylformamide or in acetonitrile with regularheating or microwave one.

Compounds of formula (VIIc) wherein X═Y═N can be prepared bynucleophilic aromatic substitution from phenol (IX) and nitro compound(VIIIc) wherein usually Z=Cl at room temperature using a base such aspotassium carbonate in a solvent such as N,N-dimethylformamide. Afurther suitable solvent is acetonitrile.

step (iii): Phenol of formula (IXa) corresponding to compound of formula(IX) wherein R₁ is H and A is the heterocycle depicted on Scheme 6 canbe prepared by an intramolecular reaction from compounds of formula (X)in presence of a catalytic amount of AuCl₃ in acetonitrile at roomtemperature or a catalytic amount of PtCl₂ in acetone with heating (asdescribed for that one in the Journal of the American Chemical Society2003, 125, 5757-5766)

step (ii): Compound of formula (X) can be prepared from compound offormula (XI) with a similar method to the one described in the Journalof the American Chemical Society 2003, 125, 5757-5766 by nucleophilicsubstitution in presence of a base such as sodium hydride in a solvente.g. DMF, with addition in a second time of an electrophile e.g.3-bromo-1-propyne.

step (ii): Phenols of formula (IXb), corresponding to compounds offormula (IX) wherein R₁ is H and A the heterocycles depicted on Scheme 7(R₆ being Me or Et), can be prepared using the corresponding compoundsof formula (XII) in presence of acidic conditions such as aqueous HCl ina solvent such as methanol.

step (i): Compounds of formula (XII) can be prepared by cyclization ofcompounds of formula (XIII) using a base such as nBuLi in a solvent suchas THF e.g. at 0° C., adding in a second time 4-methylbenzenesulfonylchloride e.g. at 0° C., then a second equivalent of a base such as nBuLie.g. from 0° C. to room temperature and stopping the reaction with adiluted protic acid such as HCl.

Optionally the two steps (i) and (ii) can be carried out in a one potfashion.

step (i): Compound of formula (XIIIa) wherein R₆=Met can be prepareddirectly from compound of formula (XV)

-   -   by lithiation using for example nBuLi in a solvent such as        hexane in presence of TMEDA from room temperature to 60° C.    -   adding in a second time acetaldehyde for example at −78° C. and        warming up the reaction mixture for example to room temperature.

step (iii): Compound of formula (XIIIb) wherein R₆=Et can be preparedfrom protected compound (XIV)

-   -   by lithiation using for example nBuLi in a solvent such as        hexane at room temperature    -   adding in a second time propanal for example at 0° C. and        warming up the reaction mixture for example to room temperature.

step (ii): Compound of formula (XIV) can be prepared from compound offormula (XV) by silylation, using for examplechloro(1,1-dimethylethyl)dimethylsilane, 1H imidazole in a solvent suchas dichloromethane at room temperature.

step (iii): phenol of the type produced by step (iii) as shown above canbe obtained from alcohol of the type produced by step (ii) as shownabove after treatment with a suitable acid such as H₂SO₄ orp-tolylsolphonic acid in a suitable solvent such as ethyl acetate,methanol or ethanol.

step (ii): The product of step (ii) as shown above can be prepared fromthe protected compound of the type produced by step (ii) as shown above

-   -   by lithiation using for example nBuLi in a solvent such as        hexane at room temperature (optionally adding CeCl₃ previously        stirred in dry THF at room temperature under argon or hydrogen        atmosphere)    -   adding in a second stage cyclobutanone for example at 0° C. and        warming up the reaction mixture for example to room temperature.

step (i): The product of step (i) as shown above can be prepared fromthe starting alcohol by silylation, using for examplechloro(1,1-dimethylethyl)dimethylsilane, 1H imidazole in a solvent suchas dichloromethane at room temperature.

step (ii): Compounds of formula (XV) can be prepared from esters offormula (XVI) using a suitable reducing agent typically LiAlH₄ in asolvent such as tetrahydrofuran at a temperature such as 0° C.

step (i): Compounds of formula (XVI) can be prepared from phenols offormula (XVII) using chloro(methyloxy)methane, a base such as DIPEA in asolvent such as dichloromethane for example from 0° C. to roomtemperature.

step (ii): The product of step (ii) as shown above can be prepared fromesters of the type produced by step (i) as shown above using a suitablereducing agent typically LiAlH₄ in a solvent such as tetrahydrofuran ata temperature such as 0° C. or room temperature.

step (i): The product of step (i) as shown above can be prepared fromthe starting phenol using chloro(methyloxy)methane, a base such as NaHin a solvent such as DMF or THF for example from 0° C. to roomtemperature.

Optionally the two steps (i) and (ii) can be carried out in a one potfashion.

step (iii): Phenols of formula (IXc), corresponding to compounds offormula (IX) wherein A is the heterocycle depicted on Scheme 10, R₁ is Hor methyl, R₇ is methyl, ethyl or isopropyl can be prepared fromcompound (XVIII) by cyclization in presence of an excess of a base suchas pyridine with heating e.g. at reflux.

step (ii): Compounds of formula (XVIII) can be prepared from compoundsof formula (XIX) by acylation for example with acetic anhydride at roomtemperature.

step (i): Compounds of formula (XIX) can be prepared from compounds offormula (XX) with hydroxylamine hydrochloride using a base such assodium acetate heating e.g. at reflux in a solvent such as a mixtureethanol/water or using pyridine as solvent and base.

Amongst the ketones of formula (XX), 1-(2,6-dihydroxyphenyl)ethanone iscommercially available e.g. from Aldrich. Ketones of formula (XXa)corresponding to ketones of formula (XX) wherein R₇=Me can be preparedfrom compounds of formula (XXI). Compounds of formula (XXI) undergoes

-   -   first a bis-acylation using e.g. acetic anhydride in presence of        a base e.g. triethylamine in a solvent e.g. dichloromethane    -   followed by a Friedel Crafts acylation with intramolecular acyl        transfer in presence of a Lewis acid such as AlCl₃ in a solvent        such as chlorobenzene with heating e.g. at 90° C.

step (iii): Ketones of formula (XXb) corresponding to ketones of formula(XX) wherein R₇=Et or iPr and R₁=H can be prepared from compounds offormula (XXII) by removal of the two protective groups under acidicconditions such as aqueous HCl solution, with heating e.g. at reflux, ina solvent such as methanol.

step (ii): Compounds of formula (XXII) can be prepared from compounds offormula (XXIII) by—lithiation with for example BuLi in a solvent such astetrahydrofuran at e.g. room temperature

-   -   adding in a second time the suitable anhydride or acyl chloride        for example at −78° C.

step (i): Compound of formula (XXIII) can be prepared from compound offormula (XXIa) corresponding to compound of formula (XXI) wherein R₁=Husing a base such as sodium hydride in a solvent such as DMF for exampleat 0° C. adding in a second time chloro(methyloxy)methane for examplefrom 0° C. to room temperature.

step (iii): Phenol of formula (IXd) corresponding to compound of formula(IX) wherein R₁ is H and A is the heterocycle depicted on Scheme 13a,can be prepared from the O-methoxy precursor (XXIV) using a reaction ofdemethylation using for example BBr₃ in a solvent such asdichloromethane at 0° C.

step (ii): Compound of formula (XXIV) can be prepared by cyclizationfrom compound of formula (XXV) using for example tributylstannane andAIBN in a solvent such as toluene e.g. at reflux.

step (i): Compound of formula (XXV) can be prepared from compound offormula (XXVI) in presence of a base such as sodium hydride in a solventsuch as DMF adding in a second time the compound of formula (XXVII)wherein Z=Cl or Br e.g. at room temperature.

Step (iv): Phenol of the type produced by step (iv) as shown above canbe prepared from TiPS protected compounds of the type produced by step(iii) as shown above removing the protective group in presence of aFluoride source such as tetrabutylammonium fluoride in a suitablesolvent such as THF at room temperature.

Step (iii): the compound of the type produced by step (iii) as shownabove can be prepared by metal-halogen exchange using butyllithium orsec-butyllithium or tert-butyllithium in a suitable solvent such as THFor Et2O or n-hexane at temperature ranging from −78° C. to roomtemperature and adding in a second stage a methylating agent such asiodomethane at temperature ranging from −78° C. to room temperature.

Step (ii): the compound of the type produced by step (ii) as shown abovecan be prepared from a protected phenol of the type produced by step (i)as shown above by using a brominating agent such as NBS in a suitablesolvent such as DMF or acetonitrile or THF at room temperature.

Step (i): The compound of the type produced by step (i) as shown abovecan be prepared from the starting phenol by silylation, using forexample chloro triisopropylsilane in presence of a base such asbutyllithium in a solvent such as THF at temperature ranging from 0° C.and room temperature.

step (iii): Compound of formula (XXVI) can be prepared from compound offormula (XXVII) by removal of the protective group under acidicconditions using for example HCl gas in a solvent such asdichloromethane.

step (ii): Compound of formula (XXVII) can be prepared from compound offormula (XXVIII) by—lithiation with for example BuLi in a solvent suchas tetrahydrofuran at e.g. −78° C.

-   -   adding in a second time a solution of iodine for example at        −70° C. and leaving the reaction at room temperature.

step (i): Compound of formula (XXVIII) can be prepared from compound offormula (XXIX) by protection of the phenol using a base e.g. sodiumhydride, in a solvent e.g. tetrahydrofuran, adding in a second timebromomethyl methyl ether e.g. at room temperature.

step (iv): Alternatively phenol of formula (IXd) corresponding tocompound of formula (IX) wherein R₁ is H and A is the heterocycledepicted on Scheme 15 can be prepared from compound of formula (XXX)using an hydroxide base for example sodium hydroxide, in a solvent suchas methanol, e.g. at room temperature.

step (iii): Compound of formula (XXX) can be prepared from compound offormula (XXXI) by cyclization with conditions presented on Scheme 13a instep (ii)

step (ii): Compound of formula (XXXI) can be prepared from compound offormula (XXXII) using a base such as potassium carbonate and anelectrophile such as 3-bromo-2-methyl-1-propene, in a solvent such asacetonitrile e.g. at room temperature.

step (i): Compound of formula (XXXII) can be prepared from compound offormula (XXXIII) by acetylation using for example acetic anhydride, abase e.g. triethylamine in a solvent e.g. dichloromethane e.g. at roomtemperature.

step (vi): Phenol of formula (IXe) corresponding to compound of formula(IX) wherein R₁ is H or methyl and A is the heterocycle depicted onScheme 16 can be prepared from compound of formula (XXXIV) by removal ofthe MOM protective group under acidic conditions using for exampleaqueous HCl in a solvent such as methanol heating e.g. at 50° C.

step (v): Compound of formula (XXXIV) can be prepared from compound offormula (XXXV) by a Mitsonobu reaction using triphenylphosine in asolvent such as tetrahydrofuran and adding diisopropyl azodicarboxylateat room temperature.

step (iv): Compound of formula (XXXV) can be prepared from compound offormula (XXXVI) in a sequential mode—deprotection in acidic conditionssuch as HCl 2N in water in ethanol

-   -   evaporation of the solvent and use of a strong base such as NaH        in a solvent such as THF at 0° C.    -   addition of MOMCl at 0° C.    -   reduction with lithium aluminium hydride at 0° C.

step (iii): Compound of formula (XXXVI) can be prepared from compound offormula (XXXVII) using a Corey-Chaykovsky cyclopropanation reactioncarried out at room temperature. To pre-form the dimethyloxosulfoniummethylide, trimethylsulfoxonium iodide can be used in presence of a basesuch as NaH in a solvent such as DMSO, the compound of formula (XXXVII),(prediluted in DMSO) being added in a second time.

step (ii): Compound of formula (XXXVII) can be prepared from compound offormula (XXXVIII) using a Wittig reaction. In order to pre-form theylide, a phosphonium salt such as methyltriphenylphosphonium bromide anda strong base such as KHMDS can be used in a solvent such as THF from 0°C. to room temperature. The compound of formula (XXXVIII) prediluted ina solvent such as THF can be added in a second time at 0° C.

step (i): Compound of formula (XXXVIII) can be prepared from compound(XXIII) by lithiation using BuLi in a solvent such as hexane at roomtemperature, this solution being added in a second time fn at −78° C. tothe electrophile e.g. ethyl chloro(oxo)acetate (prediluted e.g. in THF).

Compounds of formula (XXIII) can be prepared in a similar fashion asdescribed in Scheme 12 (step i).

step (vii): Phenol of formula (IXf) corresponding to compound of formula(IX) wherein R₁ is H and A is the heterocycle depicted on Scheme 17a canbe prepared from compound of formula (XXXIX) by removal of the MOMprotective group under acidic conditions using for example aqueous HClin a solvent such as methanol heating e.g. at 60° C.

step (vi): Compound of formula (XXXIX) can be prepared by cyclization ofcompounds of formula (XL) using a base such as triethylamine in asolvent such as THF e.g. at 0° C., adding in a second timemethanesulfonyl chloride and in a third time a strong base such aspotassium 2-methyl-2-propanolate.

step (v): Compound of formula (XL) can be prepared by hydrogenation ofthe mixture of compounds of formula (XLI) and (XL) in presence of acatalyst such as Pd/C at room temperature.

step (iv): The mixture of compounds of formula (XLI) and (XL) can beprepared from compound of formula (XLII) by reaction with a reducingagent such as lithium aluminium hydride in a solvent such as THF at 0°C.

step (iii): Compound of formula (XLII) can be prepared from phenol offormula (XLIII) using a base such as NaH and chloro(methyloxy)methane ina solvent such as DMF for example from 0° C. to room temperature.Alternatively the solvent used may be DCM and with a base such as DIPEAor TEA.

step (ii): Phenol of formula (XLIII) can be prepared from compound offormula (XLIV) by demethylation using for example boron tribromide in asolvent such as dichloromethane from 0° C. to room temperature.

step (i): Compound of formula (XLIV) can be prepared from compound offormula (XLV) by reaction with propanoic anhydride in presence of a basesuch as potassium carbonate heating e.g. at 70° C. in a solvent such asDMF followed by addition of water and heating e.g. at 120° C.

step (vii): Phenol of the type produced by step (vii) as shown abovewherein R₁ is H and A is the heterocycle depicted on Scheme 17b can beprepared from a compound of the type produced by step (vi) as shownabove by removal of the MOM protective group under acidic conditionsusing for example aqueous HCl in a solvent such as methanol heating e.g.at 60° C.

step (vi): Compound of the type produced by step (vi) as shown above canbe prepared by cyclization of compounds of the type produced by step (v)as shown above using a base such as triethylamine in a solvent such asTHF e.g. at 0° C., adding in a second time methanesulfonyl chloride andin a third time a strong base such as potassium 2-methyl-2-propanolate.

Alternatively Mitsunobu conditions can be used.

step (v): Compound of the type produced by step (v) as shown above canbe prepared by hydrogenation of the mixture of compounds of the typeproduced by step (iv) as shown above in presence of a catalyst such asPd/C at room temperature.

step (iv): The mixture of compounds of the type produced by step (iv) asshown above can be prepared from compound of the type produced by step(iii) as shown above by reaction with a reducing agent such as lithiumaluminium hydride in a solvent such as THF at 0° C.

step (iii): Compound of the type produced by step (iii) as shown abovecan be prepared from phenol of the type produced by step (ii) as shownabove using a base such as NaH and chloro(methyloxy)methane in a solventsuch as DMF or THF for example from 0° C. to room temperature.

step (ii): Phenol of the type produced by step (ii) as shown above canbe prepared from compound of the type produced by step (i) as shownabove by demethylation using for example boron tribromide in a solventsuch as dichloromethane from 0° C. to reflux.

step (i): Compound of the type produced by step (i) as shown above canbe prepared from the starting alcohol by reaction with acetic anhydridein presence of a base such as potassium carbonate heating e.g. at 70° C.in a solvent such as DMF followed by addition of water and heating e.g.at 120° C.

step (iv): Phenol of formula (IXg) corresponding to compound of formula(IX) wherein R₁ is H and A is the heterocycle depicted on Scheme 18a canbe prepared from compound (XLVI) by removal of the MOM protective groupas previously described on Scheme 16 (step vi).

step (iii): Compound of formula (XLVI) can be prepared from compound offormula (XLVII) by a Mitsonobu reaction using triphenylphosine in asolvent such as tetrahydrofuran and addingbis(1-methylethyl)(E)-1,2-diazenedicarboxylate at room temperature.

step (ii): Compound of formula (XLVII) can be prepared from compound offormula (XLVIII) by reaction with a reducing agent such as lithiumaluminium hydride in a solvent such as THF at 0° C.

step (i): Compound of formula (XLVIII) can be prepared from compound offormula (XLII) by a Corey-Chaykovsky cyclopropanation reaction, carriedout at room temperature as previously described on Scheme 16 (step(iii)).

step (iv): Phenol of the type produced by step (iv) as shown above canbe prepared from the compound the type produced by step (iii) as shownabove by removal of the MOM protective group as previously described onScheme 16 (step vi).

step (iii): Compound of the type produced by step (iii) as shown abovecan be prepared from compound of the type produced by step (ii) as shownabove by a Mitsonobu reaction using triphenylphosine in a solvent suchas tetrahydrofuran and addingbis(1-methylethyl)(E)-1,2-diazenedicarboxylate at room temperature.

step (ii): Compound of the type produced by step (ii) as shown above canbe prepared from compound of the type produced by step (i) as shownabove by reaction with a reducing agent such as lithium aluminiumhydride in a solvent such as THF at 0° C.

step (i): Compound of the type produced by step (i) as shown above canbe prepared from the starting compound by a Corey-Chaykovskycyclopropanation reaction, carried out at room temperature as previouslydescribed on Scheme 16 (step (iii)).

step (vi): Phenol of formula (IXh) corresponding to compound of formula(IX) wherein R₁ is H and A is the heterocycle depicted on Scheme 19a canbe prepared from compound (XLIX) by removal of the MOM protective groupas previously described on Scheme 16 (step vi).

step (v): Compound of formula (XLIX) can be prepared by cyclization ofcompound of formula (L) using a base such as BuLi in a solvent such ashexane e.g. at 0° C., adding in a second time 4-methylbenzenesulfonylchloride e.g. at 0° C., then in a third time a second equivalent of abase such as nBuLi e.g. at 0° C.

step (iv): Compound of formula (L) can be prepared from compound offormula (LII) sequentially—by addition a reductive agent such as lithiumaluminium hydride in a solvent such as THF e.g. at 0° C.

-   -   after work up and solvent evaporation addition of a desilylating        agent such as TBAF in a solvent such as THF.

step (iii): Compound of formula (LI) can be prepared from compound offormula (LII) by a Corey-Chaykovsky cyclopropanation reaction carriedout at room temperature as previously described on Scheme 16 (step(iii)).

step (ii): Compound of formula (LII) can be prepared from compound offormula (LIII) by olefination with for example a solution of dimethyltitanocene in a solvent such as toluene (e.g. 9% w/w) heating eg at 90°C. in a solvent such as toluene. The solution of dimethyl titanocene intoluene can be prepared by the reaction of titanocene dichloride withfor example a solution of methyllithium at −10° C.

step (i): Compound of formula (LIII) can be prepared from compound (XIV)by lithiation using BuLi in a solvent such as hexane at roomtemperature, this solution being added in a second time at −78° C. tothe electrophile e.g. ethyl chloro(oxo)acetate (prediluted e.g. in THF).

step (iv): phenol of the type produced by step (iv) as shown above canbe prepared from alcohol of the type produced by step (iii) as shownabove after treatment with a suitable acid such as H₂SO₄ orp-tolylsolphonic acid in a suitable solvent such as ethyl acetate,acetonitrile, methanol or ethanol.

step (iii): the compound of the type produced by step (iii) as shownabove can be prepared from compounds of the type produced by step (ii)as shown above by reacting with ethyl magnesium bromide from 2 to 10equivalents in a suitable solvent such as THF at temperature rangingfrom −78 C to room temperature.

step (ii): Phenol of the type produced by step (ii) as shown above canbe prepared from the compound of the type produced by step (i) as shownabove by removal of the MOM protective group using TFA in a suitablesolvent such as DCM at temperature ranging from 0° C. to roomtemperature.

step (i): the compound of the type produced by step (i) as shown abovecan be prepared from the starting compound by lithiation using BuLi in asolvent such as hexane at room temperature, this solution being added ina second time at −78° C. to the electrophile e.g. ethyl chloroformate(prediluted e.g. in THF).

Step (iv): Phenol of the type produced by step (iv) as shown above canbe prepared from a compound the type produced by step (iii) as shownabove by removal of the MOM protective group as previously described onScheme 16 (step vi).

Step (iii): the compound of the type produced by step (iii) as shownabove can be prepared by cyclopropanation (Simmons-Smith conditions) ofthe unsaturated compound of the type produced by step (ii) as shownabove using diethylzinc and diiodomethane (optionally2,4,6-trichlorophenol is added) in a solvent such as dichloromethane attemperature ranging from −40° C. to room temperature.

Step (ii): the compound of the type produced by step (ii) as shown abovecan be prepared using a wittig type reaction as described in Scheme 16(step ii).

Step (i): Compound of the type produced by step (i) as shown above canbe prepared from the starting phenol using a base such as NaH andchloro(methyloxy)methane in a solvent such as DMF or THF for examplefrom 0° C. to room temperature.

Step (ii): Phenol of the type produced by step (ii) as shown above canbe prepared from lactone of the type produced by step (i) as shown aboveusing Methylmagnesium bromide in a suitable solvent such as THF ordiethyl ether at temperature ranging from −78° C. to room temperature.Obtained compound can be treated with a suitable acid such as H2SO4 orTsOH in a suitable solvent such as ethyl acetate or acetonitrile ormethanol or ethanol.

Step (i): Lacton of the type produced by step (i) as shown above can beprepared by reaction of commercially available starting anhydride with areducing agent such as K-selectride in a solvent such as THF at 0° C.;followed by MOM-protection using a base such as DIPEA andchloro(methyloxy)methane in a solvent such as DCM for example from 0° C.to room temperature.

Step (iii): Phenol of the type produced by step (iii) as shown above canbe prepared from a boronic acid of the type produced by step (ii) asshown above using H₂O₂ and NaOH in a suitable solvent such as THF or acombination of solvents such as water/THF at room temperature.

Step (ii): the boronic acid of the type produced by step (ii) as shownabove can be prepared from boronic acid of the type produced by step (i)as shown above by lithiation using sec-BuLi in a solvent such as THF attemperature ranging from −78° C. to room temperature, followed by theaddition in a second time of a methylating agent such as iodomethane attemperature ranging from −78° C. to room temperature.

Step (i): Boronic acid of the type produced by step (i) as shown abovecan be prepared from commercially available2,2-Difluoro-1,3-benzodioxole by lithiation using sec-BuLi in a solventsuch as THF at temperature ranging from −78° C. to room temperature,followed by the addition in a second time of trimethyl borate.

Step (i): Phenol of the type produced by step (i) as shown above can beprepared from commercially available 2,2-Difluoro-1,3-benzodioxole bylithiation using sec-BuLi in a solvent such as THF at temperatureranging from −78° C. to room temperature, followed by the addition in asecond time of trimethylborate at temperature ranging from −78° C. toroom temperature and followed by the addition in a third time of H₂O₂and NaOH at room temperature.

Step (iv): Phenol of the type produced by step (iv) as shown above canbe prepared from compounds of the type produced by step (iii) as shownabove by treatment with a demethylating agent such as BBr₃ in a suitablesolvent such as dichloromethane or dichloroethane at temperature rangingfrom room temperature to reflux.

Step (iii): compound of the type produced by step (iii) as shown abovecan be prepared from compound of the type produced by step (ii) as shownabove by treatment with a suitable acid such as polyphosphoric acid neatat suitable temperature such as 110° C.

Step (ii): compound of the type produced by step (ii) as shown above canbe prepared from aminoalcohol of the type produced by step (i) as shownabove by treatment with an acylating agent such as acetyl chloride inpresence of a base such as triethylamine in a suitable solvent such asdichloromethane at a suitable temperature for example 0° C.

Step (i): Alcohol of the type produced by step (i) as shown above can beprepared from the commercially available starting ketone by treatmentwith Methyl magnesium bromide in a suitable solvent such as THF ordiethyl ether at temperature ranging from 0° C. to room temperature.

step (ii): Alternatively to the route described on Scheme 2b (step ii),compounds of formula (III) can be obtained by coupling between theaniline (IV) and the precursor (LIV) with heating e.g. at 150° C. in asolvent such as toluene.

step (i): Compounds of formula (LIV) can be prepared from N-protectedamino acid (V) and 2,2′-dithiodipyridine (LV) in presence oftriphenylphosphine at room temperature in a solvent such as THF.

step (iii): Compounds of formula (VIII) can be prepared by reaction ofureas of formula (XI) and a base such as sodium methoxide in a solventsuch as Methanol at temperature ranging from 0° C. to 60° C.

step (ii): Compounds of formula (XI) can be prepared by reaction ofcommercially available anilines of formula (IVa), wherein Z is Cl, andamino esters (hydrochloride salt) of formula (XII) in a solvent e.g.dichloromethane or ethyl acetate with a carbonylating agent e.g.triphosgene preferentially prediluted in the same solvent in presence ofa base e.g. triethylamine or diisopropylethylamine at temperatureranging from 0° C. to 60° C., optionally adding a catalytic orstechiometric amount of DMAP.

step (i): Amino esters (hydrochloride salt) of formula (XII) can beprepared from commercially available aminoacids (hydrochloride salt) offormula (XIII) by reaction with methanol in presence of a catalytic orstechiometric amount of thyonyl chloride at temperature ranging fromr.t. to reflux.

The present invention provides compounds of formula (I) or apharmaceutically acceptable salt thereof for use in therapy.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of a disease or disorderwhere a modulator of the Kv3.1 or Kv3.2 or Kv3.1 and Kv3.2 channels isrequired. As used herein, a modulator of Kv3.1 or Kv3.2 or Kv3.1 andKv3.2 is a compound which alters the properties of these channels,either positively or negatively. The altered property of the channel maybe the scale of response observed or the temporal behaviour of thechannel.

Compounds of the invention may be tested in the assay of BiologicalExample 1 to determine their modulatory properties.

Diseases or conditions that may be mediated by modulation of Kv3.1and/or Kv3.2 channels may be selected from the list below. The numbersin brackets after the listed diseases below refer to the classificationcode in Diagnostic and Statistical Manual of Mental Disorders, 4thEdition, published by the American Psychiatric Association (DSM-IV)and/or the International Classification of Diseases, 10th Edition(ICD-10).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of depression and mooddisorders including Major Depressive Episode, Manic Episode, MixedEpisode and Hypomanic Episode; Depressive Disorders including MajorDepressive Disorder, Dysthymic Disorder (300.4), Depressive Disorder NotOtherwise Specified (311); Bipolar Disorders including Bipolar IDisorder, Bipolar II Disorder (Recurrent Major Depressive Episodes withHypomanic Episodes) (296.89), Cyclothymic Disorder (301.13) and BipolarDisorder Not Otherwise Specified (296.80); Other Mood Disordersincluding Mood Disorder Due to a General Medical Condition (293.83)which includes the subtypes With Depressive Features, With MajorDepressive-like Episode, With Manic Features and With Mixed Features),Substance-Induced Mood Disorder (including the subtypes With DepressiveFeatures, With Manic Features and With Mixed Features) and Mood DisorderNot Otherwise Specified (296.90); Seasonal affective disorder.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of schizophreniaincluding the subtypes Paranoid Type (295.30), Disorganised Type(295.10), Catatonic Type (295.20), Undifferentiated Type (295.90) andResidual Type (295.60); Schizophreniform Disorder (295.40);Schizoaffective Disorder (295.70) including the subtypes Bipolar Typeand Depressive Type; Delusional Disorder (297.1) including the subtypesErotomanic Type, Grandiose Type, Jealous Type, Persecutory Type, SomaticType, Mixed Type and Unspecified Type; Brief Psychotic Disorder (298.8);Shared Psychotic Disorder (297.3); Psychotic Disorder Due to a GeneralMedical Condition including the subtypes With Delusions and WithHallucinations; Substance-Induced Psychotic Disorder including thesubtypes With Delusions (293.81) and With Hallucinations (293.82); andPsychotic Disorder Not Otherwise Specified (298.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of anxiety disordersincluding Panic Attack; Panic Disorder including Panic Disorder withoutAgoraphobia (300.01) and Panic Disorder with Agoraphobia (300.21);Agoraphobia; Agoraphobia Without History of Panic Disorder (300.22),Specific Phobia (300.29, formerly Simple Phobia) including the subtypesAnimal Type, Natural Environment Type, Blood-Injection-Injury Type,Situational Type and Other Type), Social Phobia (Social AnxietyDisorder, 300.23), Obsessive-Compulsive Disorder (300.3), PosttraumaticStress Disorder (309.81), Acute Stress Disorder (308.3), GeneralizedAnxiety Disorder (300.02), Anxiety Disorder Due to a General MedicalCondition (293.84), Substance-Induced Anxiety Disorder, SeparationAnxiety Disorder (309.21), Adjustment Disorders with Anxiety (309.24)and Anxiety Disorder Not Otherwise Specified (300.00).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of substance-relateddisorders including Substance Use Disorders such as SubstanceDependence, Substance Craving and Substance Abuse; Substance-InducedDisorders such as Substance Intoxication, Substance Withdrawal,Substance-Induced Delirium, Substance-Induced Persisting Dementia,Substance-Induced Persisting Amnestic Disorder, Substance-InducedPsychotic Disorder, Substance-Induced Mood Disorder, Substance-InducedAnxiety Disorder, Substance-Induced Sexual Dysfunction,Substance-Induced Sleep Disorder and Hallucinogen Persisting PerceptionDisorder (Flashbacks); Alcohol-Related Disorders such as AlcoholDependence (303.90), Alcohol Abuse (305.00), Alcohol Intoxication(303.00), Alcohol Withdrawal (291.81), Alcohol Intoxication Delirium,Alcohol Withdrawal Delirium, Alcohol-Induced Persisting Dementia,Alcohol-Induced Persisting Amnestic Disorder, Alcohol-Induced PsychoticDisorder, Alcohol-Induced Mood Disorder, Alcohol-Induced AnxietyDisorder, Alcohol-Induced Sexual Dysfunction, Alcohol-Induced SleepDisorder and Alcohol-Related Disorder Not Otherwise Specified (291.9);Amphetamine (or Amphetamine-Like)-Related Disorders such as AmphetamineDependence (304.40), Amphetamine Abuse (305.70), AmphetamineIntoxication (292.89), Amphetamine Withdrawal (292.0), AmphetamineIntoxication Delirium, Amphetamine Induced Psychotic Disorder,Amphetamine-Induced Mood Disorder, Amphetamine-Induced Anxiety Disorder,Amphetamine-Induced Sexual Dysfunction, Amphetamine-Induced SleepDisorder and Amphetamine-Related Disorder Not Otherwise Specified(292.9); Caffeine Related Disorders such as Caffeine Intoxication(305.90), Caffeine-Induced Anxiety Disorder, Caffeine-Induced SleepDisorder and Caffeine-Related Disorder Not Otherwise Specified (292.9);Cannabis-Related Disorders such as Cannabis Dependence (304.30),Cannabis Abuse (305.20), Cannabis Intoxication (292.89), CannabisIntoxication Delirium, Cannabis-Induced Psychotic Disorder,Cannabis-Induced Anxiety Disorder and Cannabis-Related Disorder NotOtherwise Specified (292.9); Cocaine-Related Disorders such as CocaineDependence (304.20), Cocaine Abuse (305.60), Cocaine Intoxication(292.89), Cocaine Withdrawal (292.0), Cocaine Intoxication Delirium,Cocaine-Induced Psychotic Disorder, Cocaine-Induced Mood Disorder,Cocaine-Induced Anxiety Disorder, Cocaine-Induced Sexual Dysfunction,Cocaine-Induced Sleep Disorder and Cocaine-Related Disorder NotOtherwise Specified (292.9); Hallucinogen-Related Disorders such asHallucinogen Dependence (304.50), Hallucinogen Abuse (305.30),Hallucinogen Intoxication (292.89), Hallucinogen Persisting PerceptionDisorder (Flashbacks) (292.89), Hallucinogen Intoxication Delirium,Hallucinogen-Induced Psychotic Disorder, Hallucinogen-Induced MoodDisorder, Hallucinogen-Induced Anxiety Disorder and Hallucinogen-RelatedDisorder Not Otherwise Specified (292.9); Inhalant-Related Disorderssuch as Inhalant Dependence (304.60), Inhalant Abuse (305.90), InhalantIntoxication (292.89), Inhalant Intoxication Delirium, Inhalant-InducedPersisting Dementia, Inhalant-Induced Psychotic Disorder,Inhalant-Induced Mood Disorder, Inhalant-Induced Anxiety Disorder andInhalant-Related Disorder Not Otherwise Specified (292.9);Nicotine-Related Disorders such as Nicotine Dependence (305.1), NicotineWithdrawal (292.0) and Nicotine-Related Disorder Not Otherwise Specified(292.9); Opioid-Related Disorders such as Opioid Dependence (304.00),Opioid Abuse (305.50), Opioid Intoxication (292.89), Opioid Withdrawal(292.0), Opioid Intoxication Delirium, Opioid-Induced PsychoticDisorder, Opioid-Induced Mood Disorder, Opioid-Induced SexualDysfunction, Opioid-Induced Sleep Disorder and Opioid-Related DisorderNot Otherwise Specified (292.9); Phencyclidine (orPhencyclidine-Like)-Related Disorders such as Phencyclidine Dependence(304.60), Phencyclidine Abuse (305.90), Phencyclidine Intoxication(292.89), Phencyclidine Intoxication Delirium, Phencyclidine-InducedPsychotic Disorder, Phencyclidine-Induced Mood Disorder,Phencyclidine-Induced Anxiety Disorder and Phencyclidine-RelatedDisorder Not Otherwise Specified (292.9); Sedative-, Hypnotic-, orAnxiolytic-Related Disorders such as Sedative, Hypnotic, or AnxiolyticDependence (304.10), Sedative, Hypnotic, or Anxiolytic Abuse (305.40),Sedative, Hypnotic, or Anxiolytic Intoxication (292.89), Sedative,Hypnotic, or Anxiolytic Withdrawal (292.0), Sedative, Hypnotic, orAnxiolytic Intoxication Delirium, Sedative, Hypnotic, or AnxiolyticWithdrawal Delirium, Sedative-, Hypnotic-, or Anxiolytic-PersistingDementia, Sedative-, Hypnotic-, or Anxiolytic-Persisting AmnesticDisorder, Sedative-, Hypnotic-, or Anxiolytic-Induced PsychoticDisorder, Sedative-, Hypnotic-, or Anxiolytic-Induced Mood Disorder,Sedative-, Hypnotic-, or Anxiolytic-Induced Anxiety Disorder Sedative-,Hypnotic-, or Anxiolytic-Induced Sexual Dysfunction, Sedative-,Hypnotic-, or Anxiolytic-Induced Sleep Disorder and Sedative-,Hypnotic-, or Anxiolytic-Related Disorder Not Otherwise Specified(292.9); Polysubstance-Related Disorder such as Polysubstance Dependence(304.80); and Other (or Unknown) Substance-Related Disorders such asAnabolic Steroids, Nitrate Inhalants and Nitrous Oxide.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the enhancement of cognition including the treatmentof cognition impairment in other diseases such as schizophrenia, bipolardisorder, depression, other psychiatric disorders and psychoticconditions associated with cognitive impairment, e.g. Alzheimer'sdisease.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of sleep disordersincluding primary sleep disorders such as Dyssomnias such as PrimaryInsomnia (307.42), Primary Hypersomnia (307.44), Narcolepsy (347),Breathing-Related Sleep Disorders (780.59), Circadian Rhythm SleepDisorder (307.45) and Dyssomnia Not Otherwise Specified (307.47);primary sleep disorders such as Parasomnias such as Nightmare Disorder(307.47), Sleep Terror Disorder (307.46), Sleepwalking Disorder (307.46)and Parasomnia Not Otherwise Specified (307.47); Sleep Disorders Relatedto Another Mental Disorder such as Insomnia Related to Another MentalDisorder (307.42) and Hypersomnia Related to Another Mental Disorder(307.44); Sleep Disorder Due to a General Medical Condition, inparticular sleep disturbances associated with such diseases asneurological disorders, neuropathic pain, restless leg syndrome, heartand lung diseases; and Substance-Induced Sleep Disorder including thesubtypes Insomnia Type, Hypersomnia Type, Parasomnia Type and MixedType; sleep apnea and jet-lag syndrome.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of eating disorders suchas Anorexia Nervosa (307.1) including the subtypes Restricting Type andBinge-Eating/Purging Type; Bulimia Nervosa (307.51) including thesubtypes Purging Type and Nonpurging Type; Obesity; Compulsive EatingDisorder; Binge Eating Disorder; and Eating Disorder Not OtherwiseSpecified (307.50).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Autism SpectrumDisorders including Autistic Disorder (299.00), Asperger's Disorder(299.80), Rett's Disorder (299.80), Childhood Disintegrative Disorder(299.10) and Pervasive Disorder Not Otherwise Specified (299.80,including Atypical Autism).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis ofAttention-Deficit/Hyperactivity Disorder including the subtypesAttention-Deficit/Hyperactivity Disorder Combined Type (314.01),Attention-Deficit/Hyperactivity Disorder Predominantly Inattentive Type(314.00), Attention-Deficit/Hyperactivity Disorder Hyperactive-ImpulseType (314.01) and Attention-Deficit/Hyperactivity Disorder Not OtherwiseSpecified (314.9); Hyperkinetic Disorder; Disruptive Behaviour Disorderssuch as Conduct Disorder including the subtypes childhood-onset type(321.81), Adolescent-Onset Type (312.82) and Unspecified Onset (312.89),Oppositional Defiant Disorder (313.81) and Disruptive Behaviour DisorderNot Otherwise Specified; and Tic Disorders such as Tourette's Disorder(307.23).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Personality Disordersincluding the subtypes Paranoid Personality Disorder (301.0), SchizoidPersonality Disorder (301.20), Schizotypal Personality Disorder(301.22), Antisocial Personality Disorder (301.7), BorderlinePersonality Disorder (301.83), Histrionic Personality Disorder (301.50),Narcissistic Personality Disorder (301.81), Avoidant PersonalityDisorder (301.82), Dependent Personality Disorder (301.6),Obsessive-Compulsive Personality Disorder (301.4) and PersonalityDisorder Not Otherwise Specified (301.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Sexual dysfunctionsincluding Sexual Desire Disorders such as Hypoactive Sexual DesireDisorder (302.71), and Sexual Aversion Disorder (302.79); sexual arousaldisorders such as Female Sexual Arousal Disorder (302.72) and MaleErectile Disorder (302.72); orgasmic disorders such as Female OrgasmicDisorder (302.73), Male Orgasmic Disorder (302.74) and PrematureEjaculation (302.75); sexual pain disorder such as Dyspareunia (302.76)and Vaginismus (306.51); Sexual Dysfunction Not Otherwise Specified(302.70); paraphilias such as Exhibitionism (302.4), Fetishism (302.81),Frotteurism (302.89), Pedophilia (302.2), Sexual Masochism (302.83),Sexual Sadism (302.84), Transvestic Fetishism (302.3), Voyeurism(302.82) and Paraphilia Not Otherwise Specified (302.9); gender identitydisorders such as Gender Identity Disorder in Children (302.6) andGender Identity Disorder in Adolescents or Adults (302.85); and SexualDisorder Not Otherwise Specified (302.9).

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Impulse controldisorder including: Intermittent Explosive Disorder (312.34),Kleptomania (312.32), Pathological Gambling (312.31), Pyromania(312.33), Trichotillomania (312.39), Impulse-Control Disorders NotOtherwise Specified (312.3), Binge Eating, Compulsive Buying, CompulsiveSexual Behaviour and Compulsive Hoarding.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of hearing disordersincluding auditory neuropathy, auditory processing disorder, hearingloss, which includes sudden hearing loss, noise induced hearing loss,substance-induced hearing loss, and hearing loss in adults over 60(presbycusis), and tinnitus.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Ménière's disease,disorders of balance, and disorders of the inner ear.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of hyperacusis anddisturbances of loudness perception, including Fragile-X syndrome andautism.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be of use for the treatment or prophylaxis of Epilepsy, (including,but not limited to, localization-related epilepsies, generalizedepilepsies, epilepsies with both generalized and local seizures, and thelike), seizures associated with Lennox-Gastaut syndrome, seizures as acomplication of a disease or condition (such as seizures associated withencephalopathy, phenylketonuria, juvenile Gaucher's disease, Lundborg'sprogressive myoclonic epilepsy, stroke, head trauma, stress, hormonalchanges, drug use or withdrawal, alcohol use or withdrawal, sleepdeprivation, fever, infection, and the like), essential tremor, restlesslimb syndrome, partial and generalised seizures (including tonic,clonic, tonic-clonic, atonic, myoclonic, absence seizures), secondarilygeneralized seizures, temporal lobe epilepsy, absence epilepsies(including childhood, juvenile, myoclonic, photo- and pattern-induced),severe epileptic encephalopathies (including hypoxia-related andRasmussen's syndrome), febrile convulsions, epilepsy partialis continua,progressive myoclonus epilepsies (including Unverricht-Lundborg diseaseand Lafora's disease), post-traumatic seizures/epilepsy including thoserelated to head injury, simple reflex epilepsies (includingphotosensive, somatosensory and proprioceptive, audiogenic andvestibular), metabolic disorders commonly associated with epilepsy suchas pyridoxine-dependent epilepsy, Menkes' kinky hair disease, Krabbe'sdisease, epilepsy due to alcohol and drug abuse (e.g. cocaine), corticalmalformations associated with epilepsy (e.g. double cortex syndrome orsubcortical band heterotopia), chromosomal anomolies associated withseizures or epilepsy such as Partial monosomy (150)/Angelman syndrome)and the like.

In one embodiment of the invention, there is provided a compound offormula (I) or a pharmaceutically acceptable salt thereof for thetreatment or prophylaxis of depression and mood disorders, hearingdisorders, schizopherenea, substance abuse disorders, sleep disorders orepilepsy.

In one embodiment of the invention, there is provided a compound offormula (I) or a pharmaceutically acceptable salt thereof for thetreatment or prophylaxis of bipolar disorder or mania.

The term “treatment” or “treating” as used herein includes the control,mitigation, reduction, or modulation of the disease state or itssymptoms.

The term “prophylaxis” is used herein to mean preventing symptoms of adisease or disorder in a subject or preventing recurrence of symptoms ofa disease or disorder in an afflicted subject and is not limited tocomplete prevention of an affliction.

The invention also provides a method of treating or preventing a diseaseor disorder where a modulator of Kv3 is required, for example thosediseases and disorders mentioned hereinabove, which comprisesadministering to a subject in need thereof an effective amount of acompound of formula (I) or a pharmaceutically acceptable salt thereof.

The invention also provides a compound of formula (I), or apharmaceutically acceptable salt thereof, for use in the treatment orprophylaxis of a disease or disorder where a modulator of Kv3 isrequired, for example those diseases and disorders mentionedhereinabove.

The invention also provides the use of a compound of formula (I), or apharmaceutically acceptable salt thereof, in the manufacture of amedicament for the treatment or prophylaxis of a disease or disorderwhere a modulator of Kv3 is required, for example those diseases anddisorders mentioned hereinabove.

The invention also provides a method of treating depression and mooddisorders, schizopherenea, substance abuse disorders, sleep disorders orepilepsy, for example for those indications mentioned hereinabove, whichcomprises administering to a subject in need thereof an effective amountof a Kv3 modulator or a pharmaceutically acceptable salt thereof.

For use in therapy the compounds of the invention are usuallyadministered as a pharmaceutical composition. The invention alsoprovides a pharmaceutical composition comprising a compound of formula(I), or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier.

The compounds of formula (I) or their pharmaceutically acceptable saltsmay be administered by any convenient method, e.g. by oral, parenteral,buccal, sublingual, nasal, rectal or transdermal administration, and thepharmaceutical compositions adapted accordingly.

The compounds of formula (I) or their pharmaceutically acceptable saltswhich are active when given orally can be formulated as liquids orsolids, e.g. as syrups, suspensions, emulsions, tablets, capsules orlozenges.

A liquid formulation will generally consist of a suspension or solutionof the active ingredient in a suitable liquid carrier(s) e.g. an aqueoussolvent such as water, ethanol or glycerine, or a non-aqueous solvent,such as polyethylene glycol or an oil. The formulation may also containa suspending agent, preservative, flavouring and/or colouring agent.

A composition in the form of a tablet can be prepared using any suitablepharmaceutical carrier(s) routinely used for preparing solidformulations, such as magnesium stearate, starch, lactose, sucrose andcellulose.

A composition in the form of a capsule can be prepared using routineencapsulation procedures, e.g. pellets containing the active ingredientcan be prepared using standard carriers and then filled into a hardgelatin capsule; alternatively a dispersion or suspension can beprepared using any suitable pharmaceutical carrier(s), e.g. aqueousgums, celluloses, silicates or oils and the dispersion or suspensionthen filled into a soft gelatin capsule.

Typical parenteral compositions consist of a solution or suspension ofthe active ingredient in a sterile aqueous carrier or parenterallyacceptable oil, e.g. polyethylene glycol, polyvinyl pyrrolidone,lecithin, arachis oil or sesame oil. Alternatively, the solution can belyophilised and then reconstituted with a suitable solvent just prior toadministration.

Compositions for nasal administration may conveniently be formulated asaerosols, drops, gels and powders. Aerosol formulations typicallycomprise a solution or fine suspension of the active ingredient in apharmaceutically acceptable aqueous or non-aqueous solvent and areusually presented in single or multidose quantities in sterile form in asealed container which can take the form of a cartridge or refill foruse with an atomising device. Alternatively the sealed container may bea disposable dispensing device such as a single dose nasal inhaler or anaerosol dispenser fitted with a metering valve. Where the dosage formcomprises an aerosol dispenser, it will contain a propellant which canbe a compressed gas e.g. air, or an organic propellant such as afluorochlorohydrocarbon or hydrofluorocarbon. Aerosol dosage forms canalso take the form of pump-atomisers.

Compositions suitable for buccal or sublingual administration includetablets, lozenges and pastilles where the active ingredient isformulated with a carrier such as sugar and acacia, tragacanth, orgelatin and glycerin.

Compositions for rectal administration are conveniently in the form ofsuppositories containing a conventional suppository base such as cocoabutter.

Compositions suitable for transdermal administration include ointments,gels and patches.

In one embodiment the composition is in unit dose form such as a tablet,capsule or ampoule.

The composition may contain from 0.1% to 100% by weight, for examplefrom 10 to 60% by weight, of the active material, depending on themethod of administration. The composition may contain from 0% to 99% byweight, for example 40% to 90% by weight, of the carrier, depending onthe method of administration. The composition may contain from 0.05 mgto 1000 mg, for example from 1.0 mg to 500 mg, of the active material,depending on the method of administration. The composition may containfrom 50 mg to 1000 mg, for example from 100 mg to 400 mg of the carrier,depending on the method of administration. The dose of the compound usedin the treatment of the aforementioned disorders will vary in the usualway with the seriousness of the disorders, the weight of the sufferer,and other similar factors. However, as a general guide suitable unitdoses may be 0.05 to 1000 mg, more suitably 1.0 to 500 mg, and such unitdoses may be administered more than once a day, for example two or threea day. Such therapy may extend for a number of weeks or months.

The invention provides, in a further aspect, a combination comprising acompound of formula (I) or a pharmaceutically acceptable derivativethereof together with a further therapeutic agent or agents.

The invention provides a compound of formula (I), for use in combinationwith a further therapeutic agent or agents.

When the compounds are used in combination with other therapeuticagents, the compounds may be administered either sequentially orsimultaneously by any convenient route.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical formulation and thus pharmaceuticalformulations comprising a combination as defined above together with apharmaceutically acceptable carrier or excipient comprise a furtheraspect of the invention. The individual components of such combinationsmay be administered either sequentially or simultaneously in separate orcombined pharmaceutical formulations. The individual components ofcombinations may also be administered separately, through the same ordifferent routes.

When a compound of formula (I) or a pharmaceutically acceptablederivative thereof is used in combination with a second therapeuticagent active against the same disease state the dose of each compoundmay differ from that when the compound is used alone. Appropriate doseswill be readily appreciated by those skilled in the art.

A pharmaceutical composition of the invention, which may be prepared byadmixture, suitably at ambient temperature and atmospheric pressure, isusually adapted for oral, parenteral or rectal administration and, assuch, may be in the form of tablets, capsules, oral liquid preparations,powders, granules, lozenges, reconstitutable powders, injectable orinfusible solutions or suspensions or suppositories. Orallyadministrable compositions are generally preferred.

The present invention also provides Kv3 modulators, or theirpharmaceutically acceptable salts, for use in the treatment orprophylaxis of depression and mood disorders, hearing disorders,schizopherenea, substance abuse disorders, sleep disorders or epilepsy.

In particular Kv3 modulators or their pharmaceutically acceptable saltsmay be particularly useful in the treatment or prophylaxis of depressionand mood disorders including Major Depressive Episode, Manic Episode,Mixed Episode and Hypomanic Episode; Depressive Disorders includingMajor Depressive Disorder, Dysthymic Disorder (300.4), DepressiveDisorder Not Otherwise Specified (311); Bipolar Disorders includingBipolar I Disorder, Bipolar II Disorder (Recurrent Major DepressiveEpisodes with Hypomanic Episodes) (296.89), Cyclothymic Disorder(301.13) and Bipolar Disorder Not Otherwise Specified (296.80); OtherMood Disorders including Mood Disorder Due to a General MedicalCondition (293.83) which includes the subtypes With Depressive Features,With Major Depressive-like Episode, With Manic Features and With MixedFeatures), Substance-Induced Mood Disorder (including the subtypes WithDepressive Features, With Manic Features and With Mixed Features) andMood Disorder Not Otherwise Specified (296.90), Seasonal affectivedisorder.

The invention also provides a method of treating depression and mooddisorders, hearing disorders, schizopherenea, substance abuse disorders,sleep disorders or epilepsy, including for example those disordersmentioned hereinabove, which comprises administering to a subject inneed thereof an effective amount of Kv3 modulator or a pharmaceuticallyacceptable salt thereof.

The invention also provides a Kv3 modulator, or a pharmaceuticallyacceptable salt thereof, for use in the treatment or prophylaxis ofdepression and mood disorders, hearing disorders, schizopherenea,substance abuse disorders, sleep disorders or epilepsy, including forexample those disorders mentioned hereinabove.

The invention also provides the use of a Kv3 modulator, or apharmaceutically acceptable salt thereof, in the manufacture of amedicament for the treatment or prophylaxis of depression and mooddisorders, hearing disorders, schizopherenea, substance abuse disorders,sleep disorders or epilepsy, including for example those disordersmentioned hereinabove.

For use in therapy the Kv3 modulators are usually administered as apharmaceutical composition for example a composition comprising a Kv3modulator or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier. Examples of such compositions, andmethods of administration thereof, which compositions comprise acompound of formula (I) or a pharmaceutically acceptable salt thereof,are described hereinabove. Such compositions and methods ofadministration may also be used for other Kv3 modulators orpharmaceutically acceptable salts thereof, in the treatment ofdepression and mood disorders, hearing disorders, schizopherenea,substance abuse disorders, sleep disorders or epilepsy, including forexample those disorders mentioned hereinabove.

Furthermore, the invention relates to a method for manufacturingcompounds of formula I, to novel intermediates of use in the manufactureof compounds of formula I and to the manufacture of such intermediates.

Particular intermediates of interest include:

-   3,3-dimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 50)

-   spiro[1-benzofuran-3,1′-cyclopropan]-4-ol (Intermediate 85)

-   7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol (Intermediate    156)

and

-   3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 184)

Other intermediates of interest are the anilides of formula (IV):

Especially of interest are the anilides:

wherein:

X is C or N;

Y is C or N; and

the group W is selected from:

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention is illustrated, by way of example only, withreference to the following figures in which:

FIG. 1 a hKv3.2 currents recorded using the assay described inBiological Example 1. Data shown are the individual currents over theperiod of the depolarising voltage step to −15 mV recorded from 4different cells at two concentrations of the compound of ReferenceExample RE1. The data are fitted by a single exponential curve (solidlines) using the fitting procedure in Prism version 5 (Graphpad SoftwareInc).

FIG. 1 b hKv3.2 currents recorded using the assay described inBiological Example 1. Data shown are the individual currents over theperiod of the depolarising voltage step to −15 mV recorded from 2different cells at two concentrations of compound of Reference ExampleRE3. The data are fitted by a single exponential curve (solid lines)using the fitting procedure in Prism version 5 (Graphpad Software Inc).

FIG. 2 Recordings made from identified “fast-firing” interneurons in thesomatosensory cortex of the mouse.

FIG. 3 The frequency of action potentials recorded fromparvalbumin-positive interneurons in the somatosensory cortex of themouse, evoked by depolarizing current steps

FIG. 4 The half-width of evoked action potentials fromparvalbumin-positive interneurons in the somatosensory cortex of themouse

FIG. 5 High-voltage activated potassium currents recorded from visuallyidentified MNTB neurons in the mouse, in vitro

FIG. 6 a Expression of Kv3.1b mRNA in the suprachiasmatic nucleus ofmice sacrificed during at different Circadian times over a 24-hourlight-dark cycle.

FIG. 6 b Expression of Kv3.2 mRNA in the superchiasmatic nucleus.

EXPERIMENTAL

The invention is illustrated by the Compounds described below. Thefollowing Examples describe the laboratory synthesis of specificcompounds of the invention and are not meant to limit the scope of theinvention in any way with respect to compounds or processes. It isunderstood that, although specific reagents, solvents, temperatures andtime periods are used, there are many possible equivalent alternativesthat can be used to produce similar results. This invention is meant toinclude such equivalents.

Analytical Equipment

Starting materials, reagents and solvents were obtained from commercialsuppliers and used without further purification unless otherwise stated.Unless otherwise stated, all compounds with chiral centres are racemic.Where reactions are described as having been carried out in a similarmanner to earlier, more completely described reactions, the generalreaction conditions used were essentially the same. Work up conditionsused were of the types standard in the art, but may have been adaptedfrom one reaction to another. The starting material may not necessarilyhave been prepared from the batch referred to. Compounds synthesised mayhave various purities ranging from for example 85% to 98%. Calculationsof number of moles and yield are in some cases adjusted for this.

Proton Magnetic Resonance (NMR) spectra were recorded either on Varianinstruments at 300, 400, 500 or 600 MHz, or on Bruker instruments at 400MHz. Chemical shifts are reported in ppm (δ) using the residual solventline as internal standard. Splitting patterns are designed as s(singlet), br.s (broad singlet), d (doublet), t (triplet), q (quartet),dd (doublet of doublets), dt (doublet of triplets) and m (multiplet).The NMR spectra were recorded at temperatures ranging from 25 to 30° C.

HPLC analyses indicated by HPLC (walk-up): rt=x min, were performed on aAgilent 1100 series instrument using a Luna 3u C18(2) 100A column(50×2.0 mm, 3 μm particle size) [Mobile phase, solvent A: (water+0.05%TFA), solvent B: (acetonitrile+0.05% TFA), Gradient: 100% of (A) to 95%of (B) in 8 min. Column T=40° C. Flow rate=1 ml/min. UV detectionwavelength=220 nm]. The use of this methodology is indicated by “HPLC”in the analytic characterization of the described compounds.

Direct infusion Mass spectra (MS) were run on an Agilent 1100 SeriesLC/MSD Mass Spectrometer, operating in ES (+) and ES (−) ionization mode[ES (+): Mass range: 100-1000 amu. Infusion solvent: water+0.1%HCO2H/CH3CN 50/50. ES (−): Mass range: 100-1000 amu. Infusion solvent:water+0.05% NH4OH/CH3CN 50/50]. The use of this methodology is indicatedby “MS_(—)1 (ESI)” in the analytic characterization of the describedcompounds.

Alternatively, Mass spectra (MS) were run on a mass spectrometer,operating in ES (+) and ES (−) ionization mode coupled with an HPLCinstrument Agilent 1100 Series [LC/MS-ESI(+) analyses were performed ona Supelcosil ABZ+Plus (33×4.6 mm, 3 μm) (mobile phase: from10%[CH₃CN+0.05% TFA] to 90%[CH₃CN+0.05% TFA] and 10% [water] in 2.2 min,under these conditions for 2.8 min. T=45° C., flux=0.9 mL/min)]. The useof this methodology is indicated by “MS_(—)2(ESI)” in the analyticcharacterization of the described compounds.

HPLC-Mass spectra (HPLC-MS) were taken on an Agilent 1100 Series LC/MSDMass Spectrometer coupled with HPLC instrument Agilent 1100 Series,operating in positive or negative electrospray ionization mode and inboth acidic and basic gradient conditions. Acidic gradient: LC/MS-ES (+or −) analyses were performed on a Supelcosil ABZ+Plus column (33×4.6mm, 3 μm). Mobile phase: A: (water+0.1% HCO2H)/B: CH3CN. Gradient(standard method): t=0 min 0% (B), from 0% (B) to 95% (B) in 5 minlasting for 1.5 min, from 95% (B) to 0% (B) in 0.1 min, stop time 8.5min. Column T=r.t. Flow rate=1 ml/min. The use of this methodology isindicated by “LC-MS_A” in the analytic characterization of the describedcompounds. Basic gradient: LC/MS-ES (+ or −) analyses were performed onan XTerra MS C18 column (30×4.6 mm, 2.5 μm). Mobile phase: A: (5 mM aq.NH4HCO3+ammonia (pH 10))/B: CH3CN. Gradient: t=0 min 0% (B), from 0% (B)to 50% (B) in 0.4 min, from 50% (B) to 95% (B) in 3.6 min lasting for 1min, from 95% (B) to 0% (B) in 0.1 min, stop time 5.8 min. Column T=r.t.Flow rate=1.5 mL/min]. Mass range ES (+ or −): 100-1000 amu. UVdetection range: 220-350 nm. The use of this methodology is indicated by“LC-MS_B” in the analytic characterization of the described compounds.Quality Control: LC/MS-ES+ under acidic conditions was performed on aZorbax SB C18 column (1.8 μm 3×50 mm). Mobile phase: A: (H2O+0.05% TFAby vol.)/B: (CH3CN+0.05% TFA by vol). Gradient: t=0 min 0% (B), from 0to 95% (B) in 2.5 min, 95% (B) for 0.2 min, from 95 to 100% (B) in 0.2min, 100% (B) for 0.4 min, From 100% to 0% (B) in 0.1 min. Stop time 4min. Column T=60° C. Flow rate: 1.5 ml/min. Mass range ES+: (100-1000amu, F=60). UV detection wavelengths: DAD 1A=220.8, DAD 1B=254.8. Theuse of this methodology is indicated by “LC/MS: QC_(—)3_MIN” in theanalytic characterization of the described compounds.

Ultra Performance Liquid Chromatography with an Acidic Gradient:

Total ion current (TIC) and DAD UV chromatographic traces together withMS and UV spectra associated with the peaks were taken on a UPLC/MSAcquity™ system equipped with 2996 PDA detector and coupled to a WatersMicromass ZQ™ mass spectrometer operating in positive or negativeelectrospray ionisation mode [LC/MS−ES (+ or −): analyses were performedusing an Acquity™ UPLC BEH C18 column (50×2.1 mm, 1.7 μm particle size).General Method: Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.06%HCO2H). Gradient: t=0 min 3% (B), t=0.05 min 6% (B), t=0.57 min 70% (B),t=1.06 min 99% (B) lasting for 0.389 min, t=1.45 min 3% (B), stop time1.5 min. Column T=40° C. Flow rate=1.0 mL/min. Mass range: ES (+):100-1000 amu. ES (−): 100-800 amu. UV detection range: 210-350 nm. Theuse of this methodology is indicated by “UPLC” in the analyticcharacterization of the described compounds. 1^(st) Focussed Method:Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.1% HCO2H). Gradient: t=0min 3% (B), t=1.06 min 99% (B), t=1.45 min 99% (B), t=1.46 min 3% (B),stop time 1.5 min. Column T=40° C. Flow rate=1.0 mL/min. Mass range: ES(+): 100-1000 amu. ES (−): 100-800 amu. UV detection range: 210-350 nm.The use of this methodology is indicated by “UPLC_s” in the analyticcharacterization of the described compounds. 2^(nd) Focussed Method:Mobile phase: A: (water+0.1% HCO2H)/B: (CH3CN+0.1% HCO2H). Gradient: t=0min 3% (B), t=1.5 min 100% (B), t=1.9 min 100% (B), t=2 min 3% (B), stoptime 2 min. Column T=40° C. Flow rate=1.0 mL/min. Mass range: ES (+):100-1000 amu. ES (−): 100-800 amu. UV detection range: 210-350 nm. Theuse of this methodology is indicated by “UPLC_ipqc” in the analyticcharacterization of the described compounds.

Ultra Performance Liquid Chromatography with a Basic Gradient:

Total ion current (TIC) and DAD UV chromatographic traces together withMS and UV spectra associated with the peaks were taken on a UPLC/MSAcquity™ system equipped with PDA detector and coupled to a Waters SQDmass spectrometer operating in positive and negative alternateelectrospray ionisation mode [LC/MS−ES+/−: analyses were performed usingan Acquity™ UPLC BEH C18 column (50×2.1 mm, 1.7 μm particle size).Mobile phase: A: (10 mM aqueous solution of NH4HCO3 (adjusted to pH 10with ammonia))/B: CH3CN. Gradient: t=0 min 3% (B), t=1.06 min 99% (B)lasting for 0.39 min, t=1.46 min 3% (B), stop time 1.5 min. Column T=40°C. Flow rate=1.0 mL/min. Mass range: ES (+): 100-1000 amu. ES (−):100-1000 amu. UV detection range: 220-350 nm. The use of thismethodology is indicated by “UPLC_B” in the analytic characterization ofthe described compounds.

For reactions involving microwave irradiation, a Personal ChemistryEmrys™ Optimizer was used or a Biotage Initiator

In a number of preparations, purification was performed using Biotagemanual flash chromatography (Flash+), Biotage automatic flashchromatography (Horizon, SP1 and SP4), Companion CombiFlash (ISCO)automatic flash chromatography, Flash Master Personal or Vac Mastersystems.

Flash chromatographies were carried out on silica gel 230-400 mesh(supplied by Merck AG Darmstadt, Germany) or on silica gel 300-400 mesh(supplied by Sinopharm Chemical Reagent Co., Ltd.), Varian Mega Be—Sipre-packed cartridges, pre-packed Biotage silica cartridges (e.g.Biotage SNAP cartridge), KP-NH prepacked flash cartridges, ISOLUTE NH₂prepacked cartridges or ISCO RediSep Silica cartridges.

SPE-SCX cartridges are ion exchange solid phase extraction columnssupplied by Varian. The eluent used with SPE-SCX cartridges is DCM andMeOH or only MeOH followed by ammonia solution in MeOH. The collectedfractions are those eluted with the ammonia solution in MeOH unlessotherwise stated.

SPE-Si cartridges are silica solid phase extraction columns supplied byVarian.

In a number of preparations, purification was performed on aMass-Directed Autopurification (MDAP) system FractionIynx™ equipped withWaters 2996 PDA detector and coupled with ZQ™ mass spectrometer (Waters)operating in positive and negative electrospray ionisation mode ES+, ES−(mass range 100-1000 or 100-900)

A set of semi-preparative gradients have been used:

Method A: Chromatographic Basic Conditions

-   Column: XTerra Prep MS C18 OBD (150 mm×30 mm 10 μm particle size) at    room temperature-   Mobile phase: A: (water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia)), B: acetonitrile-   Flow rate: 40 ml/min-   Gradient: 10% (B) for 0.5 min, from 10% (B) to 95% (B) in 12.5 min,    from 95% (B) to 100% (B) in 3 min

Method B: Chromatographic Basic Conditions

-   Column: XTerra Prep MS C18 OBD (150 mm×30 mm 10 μm particle size) at    room temperature-   Mobile phase: A: water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia), B: acetonitrile-   Flow rate: 40 ml/min-   Gradient: from 20% to 25% (B) in 1 min, from 25% (B) to 65% (B) in    12 min, from 65% (B) to 100% (B) in 0.5 min

Method C: Chromatographic Basic Conditions

-   Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at    room temperature-   Mobile phase: A: water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia), B: acetonitrile-   Flow rate: 17 ml/min-   Gradient: from 20% (B) to 25% (B) in 1 min, from 25% (B) to 55% (B)    in 9 min, from 55% (B) to 100% (B) in 2 min, return to 20% (B) in    0.1 min

Method D: Chromatographic Acidic Conditions

-   Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at    room temperature-   Mobile phase: A: (water+0.1% formic acid in water); B: acetonitrile-   Flow rate: 17 ml/min-   Gradient: from 20% (B) to 25% B in 1 min, from 25% (B) to 55% (B) in    9 min, from 55% (B) to 100% (B) in 2 min, return to 20% (B) in 0.1    min

Method E: Chromatographic Basic Conditions

-   Column: Waters Xbridge C18 OBD (50 mm×19 mm 5 μm particle size) at    room temperature-   Mobile phase: A: (water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia)), B: acetonitrile-   Flow rate: 17 ml/min-   Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 70% (B)    in 7 min, from 70% (B) to 100% (B) in 1 min, 100% (B) for 2 min,    return to 10% (B) in 0.1 min

Method F: Chromatographic Basic Conditions

-   Column: Phenomenex Gemini AXIA C18 (50×21.2 mm 5 μm particle size)-   Mobile phase: A: water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia), B: acetonitrile-   Flow rate: 17 ml/min-   Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 65% (B)    in 8 min, from 65% (B) to 100% (B) in 1 min, return to 10% (B) in 1    min.

Method G: Chromatographic Basic Conditions

-   Column: Phenomenex Gemini AXIA C18 (50×21.2 mm 5 μm particle size)-   Mobile phase: A: water+10 mM aqueous solution of ammonium    bicarbonate (adjusted to pH 10 with ammonia), B: acetonitrile-   Flow rate: 17 ml/min-   Gradient: from 10% (B) to 15% (B) in 1 min, from 15% (B) to 70% (B)    in 7 min, from 70% (B) to 100% (B) in 1 min, 100% (B) during 2 min,    return to 10% (B) in 0.1 min.

ABBREVIATIONS

-   CDCl₃ deutrated chloroform-   cHex cyclohexane-   CV column volume-   DCM dichloromethane-   DIPEA N,N-diisopropylethylamine-   DMAP 4-dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO dimethylsulfoxide-   DMSO-d₆ deutrated dimethylsulfoxide-   EDC.HCl N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride-   Et₂O diethyl ether-   EtOAc ethyl acetate-   h hours-   H₂ gaseous hydrogen-   HATU    (O-7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro    phosphate)-   HBTU O-benzotriazol-1-yl-tetramethyluronium hexafluorophosphate-   HCO2H formic acid-   HCl hydrogen chloride-   HNO₃ nitric acid-   HOBt.H₂O 1-hydroxybenzyltriazole hydrate-   H₂SO₄ sulfuric acid-   K₂CO₃ potassium carbonate-   KHDMS potassium hexamethyldisilazide-   KOH potassium hydroxide-   MeCN/CH₃CN acetonitrile-   MeOH methanol-   MeOD deutrated methanol-   MDAP mass-directed autopurification-   MOM methoxymethyl-   N₂ gaseous nitrogen-   NaBH(OAc)₃ sodium triacethoxyborohydride-   NaHCO₃ sodium hydrogenocarbonate-   NaNO₂ sodium nitrite-   Na₂CO₃ sodium carbonate-   NaOH sodium hydroxide-   NH4OH ammonium hydroxide-   NH4HCO3H ammonium bicarbonate-   NMR Nuclear Magnetic Resonance-   Pd/C palladium on charcoal-   PE petroleum ether-   r.t. room temperature-   SCRC Sinopharm Chemical Reagent Co., Ltd-   T3P Propylphosphonic anhydride-   tBuOK potassium tert-butoxide-   TBTU o-Benzotriazol-1-yl-n,n,n′,n′-tetramethyluronium    tetrafluoroborate-   TEA triethylamine-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TsOH*H₂O 4-methylbenzenesulfonic acid hydrate, p-toluenesulfonic    acid hydrate

Intermediate 1 2-[(2-propyn-1-yloxy)methyl]furan

To a suspension of sodium hydride (1.570 g, 39.2 mmol) in DMF (46 ml)stirred under argon at 0° C. was dropped a solution of 2-furanylmethanol(3.5 g, 35.7 mmol) in DMF (4 ml) in 20 minutes. The reaction mixture wasstirred at 0° C. for 15 minutes. 3-bromo-1-propyne (4.24 g, 35.7 mmol)80% in toluene was dropped in 10 minutes at 0° C., then the mixture wasleft stirring at room temperature overnight. Water was added and thenthe mixture was extracted with ethyl ether 3 times. The organic phasewas dried over sodium sulphate and concentrated under vacuum. Theresidue was purified by flash chromatography on silica (Biotage SP1instrument), eluting with a gradient cyclohexane/ethyl acetate 95/5 to85/15. Evaporation afforded the title compound (1.63 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.66 (1H, d), 6.41-6.49 (2H, m), 4.46(2H, s), 4.12 (2H, d), 3.48 (1H, t); UPLC-MS: 0.66 min

Intermediate 2-3 1,3-dihydro-2-benzofuran-4-ol and1,3-dihydro-2-benzofuran-5-ol

To a solution of [2-[(2-propyn-1-yloxy)methyl]furan] (Intermediate 1,1.63 g) in acetonitrile (60 ml) stirred under argon at room temperaturewas added neat gold trichloride (0.182 g, 0.599 mmol). The reactionmixture was stirred overnight at room temperature. Gold trichloride wasthen added (120 mg) and after 2 hours another gold trichloride additionwas carried out (226 mg). After 1 hour the mixture was concentrated andthe crude was purified by flash chromatography (Biotage SP1), elutingwith cyclohexane/ethyl acetate 90/10. Evaporation of the two fractionsgave respectively the title compounds: 1,3-dihydro-2-benzofuran-4-ol(100 mg) and 1,3-dihydro-2-benzofuran-5-ol (356 mg)

Intermediate 2: 1,3-dihydro-2-benzofuran-4-ol: ¹H NMR (400 MHz,DMSO-d₆): δ ppm 9.59 (1H, s), 7.15-7.00 (1H, m), 6.78-6.60 (2H, m),5.03-4.84 (4H, m); UPLC-MS: 0.41 min, 135 [M−H]−

Intermediate 3: 1,3-dihydro-2-benzofuran-5-ol: ¹H NMR (400 MHz,DMSO-d₆): δ ppm 9.36 (1H, s), 7.11-7.02 (1H, m), 6.70-6.61 (2H, m), 4.89(4H, m); UPLC-MS: 0.42 min, 135 [M−H]−

Intermediate 4 4-[(4-nitrophenyl)oxy]-1,3-dihydro-2-benzofuran

A suspension of potassium carbonate (670 mg, 4.85 mmol),1,3-dihydro-2-benzofuran-4-ol (Intermediate 2, 110 mg) and1-fluoro-4-nitrobenzene (114 mg, 0.808 mmol) in N,N-dimethylformamide(DMF) (5 ml) was heated under microwave irradiation at 100° C. for 3×30minutes. The mixture was concentrated. 2 ml of water were added and thendichloromethane. The phases were separated and the aqueous phase wasextracted twice with dichloromethane. The organic phase was dried oversodium sulphate and concentrated under vacuum to afford the titlecompound, which was directly used in the next step.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.30-8.20 (2H, m), 7.46-7.36 (1H, m),7.32-7.22 (m, 1H), 7.18-7.10 (2H, m), 7.09-7.04 (1H, m), 5.14-4.79 (4H,m); UPLC-MS: 0.98 min

Intermediate 5 4-(1,3-dihydro-2-benzofuran-4-yloxy)aniline

A solution of 4-[(4-nitrophenyl)oxy]-1,3-dihydro-2-benzofuran(Intermediate 4, 208 mg), hydrazine hydrate (0.051 ml, 1.618 mmol) andPd/C (172 mg, 0.162 mmol) in ethanol (6 ml) was stirred under argon at90° C. After 1.5 hour the mixture was cooled to room temperature andthen filtered over celite. The celite was washed with methanol. Theorganic phase was concentrated to afford title compound (136 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.20 (1H, s), 6.96 (1H, d), 6.79-6.73(2H, m), 6.62-6.55 (3H, m), 5.00 (4H, s), 4.88 (2H, s); UPLC-MS: 0.72min, 228 [M+1]+

Intermediate 61,1-dimethylethyl((1R)-2-{[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]amino}-1-methyl-2-oxoethyl)carbamate

A suspension of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (97 mg,0.515 mmol), DIPEA (0.138 ml, 0.792 mmol) and TBTU (191 mg, 0.594 mmol)in 1,2-dichloroethane (3 ml) was stirred under argon at room temperaturefor 45 min. 4-(1,3-dihydro-2-benzofuran-4-yloxy)aniline (Intermediate 5,90 mg) was added and the mixture was left under stirring at roomtemperature overnight. Brine was added and the mixture was separated ina separator tube. The aqueous phase was extracted twice withdichloromethane. The organic phase was dried over sodium sulphate andconcentrated under vacuum. The residue was purified by chromatography(Biotage SP1), using as eluents a gradient Cyclohexane/Ethyl acetatefrom 100:0 to 85:15 to afford the title compound (109 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.97 (1H, m), 7.67-7.57 (2H, m),7.32-7.23 (1H, m), 7.11-7.04 (2H, m), 7.04-6.96 (2H, m), 6.79-6.71 (1H,m), 5.08-4.99 (2H, m), 4.93-4.83 (2H, m), 4.15-4.05 (1H, m), 1.39 (9H,s), 1.29-1.22 (3H, m); UPLC-MS: 0.85 min, 399 [M+H]+

Intermediate 7N¹-[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]-D-alaninamide

A solution of1,1-dimethylethyl((1R)-2-{[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]amino}-1-methyl-2-oxoethyl)carbamate(Intermediate 6, 108 mg) and TFA (1 ml) in dichloromethane (4 ml) wasstirred under argon at room temperature. The reaction mixture wasstirred at room temperature for 1 hour. The mixture was concentrated andthe residue was purified by SCX to afford the title compound (81 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm: 7.66 (2H, d), 7.31-7.24 (1H, m),7.09-7.04 (1H, m), 6.99 (2H, d), 6.71-6.77 (1H, m), 5.03 (2H, s), 4.87(2H, s), 3.40-3.47 (1H, m), 1.21 (3H, d); UPLC-MS: 0.67 min, 299 [M+H]+

Intermediate 8 1-(2,6-dihydroxyphenyl)ethanone oxime

A solution of hydroxylamine hydrochloride (756 mg, 10.9 mmol) and sodiumacetate trihydrate (1.71 g, 12.6 mmol) dissolved in 23 ml of a mixtureof EtOH/H2O (7/3) was added to a solution of 2′,6′-dihydroxyacetophenone(1.5 g, 9.9 mmol) in 12 ml of a 7/3 EtOH/H2O mixture. After refluxingand stirring under N2 for 2 hours, additional hydroxylaminehydrochloride (340 mg, 4.9 mmol) and sodium acetate trihydrate (570 mg,4.19 mmol) dissolved in 7 ml of water were added and the reflux wascontinued for additional 30 minutes. After cooling down to roomtemperature, the volatiles were removed. Then water was added and thesolid obtained was filtered, washed with water and dried. This afforded1.3 g of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.99 (1H, br. s), 9.63 (2H, br. s),7.01-6.81 (1H, m), 6.44-6.19 (2H, m), 2.11 (3H, s).

Intermediate 9 1-(2,6-dihydroxyphenyl)ethanone O-acetyloxime

To 1-(2,6-dihydroxyphenyl)ethanone oxime (Intermediate 8, 588 mg),acetic anhydride (1.97 ml, 20.8 mmol) was added and the reaction mixturewas stirred at room temperature for 15 minutes. After the removal of thevolatiles, water was added and the solid obtained was filtered and driedunder high vacuum affording 437 mg of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.62 (2H, br. s), 7.07-6.96 (1H, m),6.37 (2H, d), 2.19 (6H, s).

Intermediate 10 3-methyl-1,2-benzisoxazol-4-ol

To 1-(2,6-dihydroxyphenyl)ethanone O-acetyloxime (Intermediate 9, 437mg) pyridine (4.0 ml) was added and the reaction mixture was stirred atreflux for 2 hours. After the addition of HCl (4.0 ml of a 5M aqueoussolution), the mixture was extracted 3 times with Et2O and the collectedorganic layers were washed with HCl (1M, aqueous solution). The organicphase was dried over sodium sulphate and filtered. Evaporation affordedthe title compound (137 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.67 (1H, br. s), 7.48-7.33 (1H, m),7.08-6.99 (1H, m), 6.69-6.62 (1H, m), 2.58 (3H, s).

Intermediate 11 3-methyl-4-[(4-nitrophenyl)oxy]-1,2-benzisoxazole

3-methyl-1,2-benzisoxazol-4-ol (Intermediate 10, 137 mg) was stirredwith 1-fluoro-4-nitrobenzene (130 mg, 0.92 mmol) in DMF (3.0 ml) withpotassium carbonate (381 mg, 2.8 mmol). The reaction mixture was heatedat 110° C. for 1 hour under microwave irradiation After removal of thevolatiles, the residue was purified by silica gel chromatography elutingwith CyHex/EtOAc (100%/0% to 0%/100%) to afford the title compound (100mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.41-8.25 (2H, m), 7.78-7.57 (2H, m),7.35-7.24 (2H, m), 7.09-6.98 (1H, m), 2.45 (3H, s); UPLC-MS: 0.83 min,271 [M+H]+.

Intermediate 12 4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]aniline

3-methyl-4-[(4-nitrophenyl)oxy]-1,2-benzisoxazole (Intermediate 11, 100mg) was dissolved under nitrogen atmosphere in 5.0 ml of EtOH. Tin(II)chloride dihydrate (417 mg, 1.85 mmol) was added. The reaction mixturewas then stirred at 90° C. for 5 hours. After removal of the volatiles,water was added and the reaction mixture was extracted two times withethyl acetate. The collected organic layers were washed with a 5%aqueous solution of NaHCO3, dried over sodium sulphate, filtered andevaporated. The crude obtained was purified by a NH column and elutedwith DCM/MeOH (100/0, then a gradient from 100/0 to 90/10, then 90/10)to afford 30 mg of the title compound.

UPLC-MS: 0.63 min, 241 [M+H]+

Intermediate 131,1-dimethylethyl[(1R)-1-methyl-2-({4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-2-oxoethyl]carbamate

{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amine (Intermediate 12,982 mg) was dissolved in 12.0 ml of DMF. DIPEA (1.07 ml, 6.1 mmol) andHATU (1865 mg, 4.9 mmol) were added. After stirring for 15 minutes,N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (928 mg, 4.9 mmol) wasadded and the reaction mixture was stirred at 60° C. for 2 hours. Afterremoval of the volatiles, the residue was purified by silica gelchromatography eluting with a gradient CyHex/EtOAc from 100/0% to0/100%. This afforded the title compound (587 mg).

UPLC-MS_B: 0.89 min, 412 [M+H]+.

Intermediate 14N¹-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-D-alaninamide

1,1-dimethylethyl[(1R)-1-methyl-2-({4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-2-oxoethyl]carbamate(Intermediate 13, 587 mg) was dissolved in 10.0 ml of DCM and then TFA(5.0 ml) was added. The reaction mixture was stirred at room temperaturefor 1 hour. After the removal of the volatiles, the residue was purifiedwith an SCX cartridge and eluted with DCM/MeOH/NH3 (2.0 M solution inMeOH). Evaporation afforded 337 mg of the title compound.

UPLC-MS_B: 0.71 min, 312 [M+H]+.

Intermediate 15 5-methylbenzene-1,3-diyl diacetate

5-methyl-1,3-benzenediol (2.0 g, 16.11 mmol) was dissolved in 20.0 ml ofdichloromethane and TEA (11.23 ml, 81.0 mmol) was added. Then, at 0° C.,acetic anhydride (4.56 ml, 48.30 mmol) was added and the reactionmixture was stirred at room temperature for 50 hours. After the additionof water (20.0 ml) the reaction mixture was stirred for 3 hours. Thephases were then separated and the aqueous one was extracted withdichloromethane (2 times). The gathered organic phases were washed withbrine, dried over sodium sulphate, filtered and evaporated to afford thetitle compound (3.33 g).

¹H-NMR (400 MHz, CDCl₃): δ ppm 6.83 (2H, br. s), 6.75 (1H, br. s), 2.39(3H, s), 2.31 (6H, s); UPLC-MS: 0.67 min, 209 [M+H]+.

Intermediate 16 1-(2,6-dihydroxy-4-methylphenyl)ethanone

A solution of 5-methyl benzene-1,3-diyl diacetate (Intermediate 15, 3.33g) in chlorobenzene (5.0 ml) was added dropwise to a suspension of AlCl₃(6.40 g, 48.0 mmol) in chlorobenzene (15.0 ml). The reaction mixture wasstirred at 90° C. for 1 hour, then it was cooled down to roomtemperature and pipetted onto a mixture of ice and 2 M HCl aqueoussolution (16 ml). Ethyl acetate was added, the two phases wereseparated. The organic one was washed 2 times with brine, then driedover sodium sulphate, filtered and evaporated. The residue obtained waspurified by silica gel chromatography eluting with a gradientCy-Hex/EtOAc from 100/0 to 70/30 followed by an isochratic 70/30,another gradient from 70/30 to 50/50 and another isochratic 50/50. Thisafforded the title compound (940 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 11.89 (2H, s), 6.22 (2H, s), 2.63 (3H,s), 2.19 (3H, s); UPLC-MS: 0.62 min, 167 [M+H]+.

Intermediate 17 1-(2,6-dihydroxy-4-methylphenyl)ethanone oxime

A solution of hydroxylamine hydrochloride (470 mg, 6.76 mmol) and sodiumacetate trihydrate (590 mg, 4.34 mmol) dissolved in 20 ml of a mixtureof EtOH/H2O (7/3) was added to a solution of1-(2,6-dihydroxy-4-methylphenyl)ethanone (Intermediate 16, 940 mg) in 15ml of a mixture of EtOH/H2O (7/3). After refluxing and stirring under N2for 2 hours, additional hydroxylamine hydrochloride (159 mg, 2.29 mmol)and sodium acetate trihydrate (199 mg) dissolved in 5 ml of water wereadded. The reaction mixture was heated at reflux overnight. Aftercooling down to room temperature, the volatiles were removed. Water wasadded and the solid afforded was filtered, washed with water and driedgiving 829 mg of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.96 (1H, s), 9.80 (2H, s), 6.16 (2H,s), 2.13 (6H, br. s).

Intermediate 18 1-(2,6-dihydroxy-4-methylphenyl)ethanone O-acetyloxime

To 1-(2,6-dihydroxy-4-methylphenyl)ethanone oxime (Intermediate 17, 829mg), acetic anhydride (2.6 ml, 27.5 mmol) was added and the reactionmixture was stirred at room temperature for 1 hour. After the removal ofthe volatiles, the residue was washed with water, filtered and dried.This afforded 1.0 g of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.58 (2H, s), 6.20 (2H, s), 2.21-2.18(6H, m), 2.16 (3H, s); acid UPLC-MS: 0.58 min, 224 [M+H]+.

Intermediate 19 3,6-dimethyl-1,2-benzisoxazol-4-ol

To 1-(2,6-dihydroxy-4-methylphenyl)ethanone O-acetyloxime (Intermediate18, 1.0 g) pyridine (10 ml) was added and the reaction mixture wasstirred at reflux under N2 for 2 hours. After the addition of HCl (10.0ml of a 5M aqueous solution), the mixture was extracted 3 times withEt2O and the gathered organic phases were washed with HCl (1M). Theseparated organic phase was then dried over sodium sulphate, filteredand evaporated to afford 345 mg of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ 10.55 (1H, s), 6.85 (1H, bs), 6.47 (1H,bs), 2.53 (3H, s), 2.35 (3H, s).

Intermediate 20 3,6-dimethyl-4-[(4-nitrophenyl)oxy]-1,2-benzisoxazole

3,6-dimethyl-1,2-benzisoxazol-4-ol (Intermediate 19, 345 mg) wasdissolved in acetonitrile (10.0 ml) and then 1-fluoro-4-nitrobenzene(298 mg, 2.11 mmol) and potassium carbonate (877 mg, 6.34 mmol) wereadded. The reaction mixture was stirred and heated at reflux overnight.After removal of the volatiles, the residue was purified by silica gelchromatography eluting with a gradient cHex/EtOAc from 100/0 to 50/50 toafford the title compound (208 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.36-8.26 (2H, m), 7.46 (1H, s),7.33-7.26 (2H, m), 6.91 (1H, s), 2.47 (3H, s), 2.39 (3H, s); UPLC-MS:0.84 min, 285 [M+H]+.

Intermediate 21 {4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amine

3,6-dimethyl-4-[(4-nitrophenyl)oxy]-1,2-benzisoxazole (Intermediate 20,208 mg) was dissolved in ethanol (10.0 ml) and tin chloride dihydrate(991 mg, 4.39 mmol) was added. The reaction mixture was stirred andheated at reflux for 4 hours. After the removal of the volatiles, waterwas added and the reaction mixture was extracted two times with ethylacetate. The collected organic were dried over sodium sulphate, filteredand evaporated to afford 260 mg of the title compound.

UPLC-MS: 0.60 min, 255 [M+H]+.

Intermediate 221,1-dimethylethyl[(1R)-2-({4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-1-methyl-2-oxoethyl]carbamate)

{4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amine (Intermediate21, 260 mg) was dissolved in 8.0 ml of DMF. DIPEA (0.188 ml, 1.074 mmol)and HATU (327 mg, 0.86 mmol) were added. After stirring for 15 minutes,N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (163 mg, 0.86 mmol) wasadded and the reaction mixture was stirred at 60° C. for 2 hours. Afterremoval of the volatiles, the residue was purified by silica gelchromatography eluting with a gradient cHex/EtOAc from 100/0 to 0/100 toafford the title compound (74 mg).

UPLC-MS_B: 0.93 min, 426 [M+H]+.

Intermediate 23N¹-{4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-D-alaninamide

1,1-dimethylethyl[(1R)-2-({4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-1-methyl-2-oxoethyl]carbamate)(Intermediate 22, 74 mg) was dissolved in 3.0 ml of dichloromethane. TFA(1.5 ml) was added and the reaction mixture was stirred at roomtemperature for 1 hour. After removal of the volatiles, the residue waspurified with a SCX cartridge and eluted with DCM/MeOH/NH3 (2.0 Msolution in MeOH). The evaporation of the volatiles, afforded 54 mg ofthe title compound.

UPLC-MS_B: 0.76 min, 326 [M+H]+.

Intermediate 241,1-dimethylethyl[1,1-dimethyl-2-({4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-2-oxoethyl]carbamate

{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amine (Intermediate 23,30 mg) was dissolved in 5.0 ml of DMF. DIPEA (0.033 ml, 0.19 mmol) andHATU (57 mg, 0.15 mmol) were added. After stirring for 15 minutes,N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (30.5 mg, 0.15mmol) was added and the reaction mixture was stirred at 60° C. for 2hours. After the removal of the volatiles, the residue was purified bysilica gel chromatography eluting with a gradient cHex/EtOAc (from 100/0to 0/100) to afford the title compound (34 mg).

UPLC-MS_B: 0.91 min, 426 [M+H]+.

Intermediate 252-methyl-N¹-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}alaninamide

1,1-dimethylethyl[1,1-dimethyl-2-({4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}amino)-2-oxoethyl]carbamate(Intermediate 24, 34 mg) was dissolved in 4.0 ml of dichloromethane andthen TFA (1.0 ml) was added. The reaction mixture was stirred at roomtemperature for 1 hour. After removal of the volatiles, the residue wascharged on a SCX cartridge and eluted successively with dichloromethane,MeOH, NH3 (2.0 M solution in MeOH). Evaporation afforded 18 mg of thetitle compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.84-7.70 (2H, m), 7.60-7.49 (1H, m),7.45-7.36 (1H, m), 7.21-7.13 (2H, m), 6.61-6.51 (1H, m), 2.60 (3H, s),1.33 (6H, s); basic UPLC-MS: 0.79 min, 326 [M+H]+.

Intermediate 26 1,3-bis{[(methyloxy)methyl]oxy}benzene

Resorcinol (3.0 g, 27.2 mmol) was dissolved in DMF (50.0 ml) and, at 0°C., NaH (4.36 g, 109 mmol, 60% Wt) was added. After stirring at thattemperature for 30 minutes, chloromethyl methyl ether (8.28 ml, 109mmol) was added and the reaction was allowed to reach room temperatureand was stirred overnight. The mixture was quenched with a saturatedaqueous solution of NaHCO3 and extracted with ethyl acetate. Thecollected organic layers were then washed with brine, dried, filteredand evaporated. The residue obtained was charged on a silica gel columnand eluted with Cyclohexane/EtOAc (from 100/0 to 50/50) to afford thetitle compound (4.94 g).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.26-7.15 (1H, m), 6.71-6.65 (3H, m),5.18 (4H, s) 3.39 (6H, s).

Intermediate 27 1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-1-propanone

1,3-bis{[(methyloxy)methyl]oxy}benzene (Intermediate 26, 2.02 g) wasdissolved under nitrogen in THF (7.0 mL) and a solution of BuLi (7.64 mlof a 1.6 M solution in hexane, 12.2 mmol) was added. After stirring atroom temperature for 1 hour and cooling down at −78° C., propanoicanhydride (5.23 ml, 40.8 mmol) was added and the mixture was stirred atthat temperature for 15 minutes. After quenching with water, the mixturewas extracted with ethyl acetate. The collected organic layers weredried over sodium sulphate, filtered and evaporated. The crude affordedwas then charged on a silica gel column and eluted withcyclohexane/EtOAc (from 10/0 to 9/1, then 9/1, then from 9/1 to 8/2,then 8/2) to afford the title compound (1.706 g).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.32-7.23 (1H, m), 6.81 (2H, d), 5.18(4H, s) 3.34 (6H, s), 2.71 (2H, q), 1.06 (3H, t).

Intermediate 28 1-(2,6-dihydroxyphenyl)-1-propanone

1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-1-propanone (Intermediate 27,1.7 g) was dissolved in methanol (50.0 ml) and an aqueous solution ofHCl (26.7 ml, 53.5 mmol of a 2M aqueous solution) was added. Thereaction mixture was refluxed for 2 hours. After quenching with water,the reaction mixture was extracted with ethyl acetate. The gatheredorganic layers were dried over sodium sulphate, filtered and evaporatedto afford the title compound (1.11 g).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 11.59 (2H, br. s), 7.25-7.17 (1H, m),6.37 (2H, d), 3.06 (2H, q), 1.07 (3H, t).

Intermediate 29 1-(2,6-dihydroxyphenyl)-1-propanone oxime

A solution of hydroxylamine hydrochloride (510 mg, 7.3 mmol) and sodiumacetate trihydrate (629 mg, 4.6 mmol) dissolved in 20 ml of a mixtureEtOH/H₂O (7/3) was added to a solution of1-(2,6-dihydroxyphenyl)-1-propanone (Intermediate 28, 1.0 g) in 15 ml ofa mixture EtOH/H₂O (7/3). After refluxing and stirring under N2overnight, additional hydroxylamine hydrochloride (510 mg, 7.3 mmol) andsodium acetate trihydrate (629 mg, 4.6 mmol) dissolved in 4.5 mL ofwater were added. The reaction mixture was heated at reflux anadditional 3 hours. After cooling down to room temperature, thevolatiles were removed. Then water was added and reaction mixture wasextracted with ethyl acetate. The gathered organic layers were driedover sodium sulphate, filtered and evaporated to afford the titlecompound (1.05 g).

UPLC: 0.41 min, 182 [M+H]+; 0.47 min, 182 [M+H]+.

Intermediate 30 1-(2,6-dihydroxyphenyl)-1-propanone O-acetyloxime

1-(2,6-dihydroxyphenyl)-1-propanone oxime (Intermediate 29, 716 mg) wasstirred in acetic anhydride (2.24 ml, 23.7 mmol) at room temperature for1 hour. After removal of the volatiles, the residue was washed withwater, filtered and dried. The crude compound was charged on a silicagel column (Biotage SP1 system) and eluted with Cyclohexane/EtOAc (from100/0 to all 0/100) to afford the title compound (269 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.49 (2H, br. s), 7.01-6.91 (1H, m),6.32 (2H, d), 2.54 (2H, q), 1.97 (3H, s), 1.02 (3H, t).

Intermediate 31 3-ethyl-1,2-benzisoxazol-4-ol and2-ethyl-1,3-benzoxazol-4-ol

1-(2,6-dihydroxyphenyl)-1-propanone O-acetyloxime (Intermediate 30, 269mg) was dissolved in pyridine (10.0 ml) and stirred at reflux for 2days. After the addition of an aqueous solution of HCl (5M), thereaction mixture was extracted 3 times with ethyl acetate and thegathered organic layers were washed with an aqueous solution of HCl(1M). The organic phase was dried over sodium sulphate, filtered andevaporated. The residue obtained was charged on a silica gel column(Biotage SP1 system) and eluted with cyclohexane/EtOAc (from 1/0 to 1/1,then 1/1, then from 1/1 to 0/1) to afford the title compounds as mixtureof 3-ethyl-1,2-benzisoxazol-4-ol and 2-ethyl-1,3-benzoxazol-4-ol (38 mg)

UPLC: 0.59 min, 164 [M+H]+; 0.61 min, 164 [M+H]+.

Intermediate 32 3-ethyl-4-[(5-nitro-2-pyridinyl)oxy]-1,2-benzisoxazole

A mixture of 3-ethyl-1,2-benzisoxazol-4-ol and2-ethyl-1,3-benzoxazol-4-ol (Intermediate 31, 38 mg), was dissolved inDMF (3.0 mL) and 2-chloro-5-nitropyridine (36.9 mg, 0.23 mmol) wasadded, followed by potassium carbonate (97 mg, 0.70 mmol). The reactionmixture was heated under microwave irradiation at 110° C. for 1 hour.After removal of the volatiles, the crude was charged on a silica gelcolumn and eluted with Cyclohexane/EtOAc (from 1/0 to 1/1) to afford thetitle compound (9.5 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.06-9.01 (1H, m), 8.75-8.68 (1H, m),7.76-7.65 (2H, m), 7.47 (1H, d), 7.27-7.22 (1H, m), 2.73-2.67 (2H, m),1.20 (3H, t). UPLC: 0.77 min, 286 [M+H]+.

Intermediate 33 6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinamine

3-ethyl-4-[(5-nitro-2-pyridinyl)oxy]-1,2-benzisoxazole (Intermediate 32,9 mg) was dissolved in 3.0 ml of ethanol. Then tin(II) chloridedihydrate (21.4 mg, 0.095 mmol) was added and the reaction mixture wasstirred at 80° C. for 3 hours. After quenching with water and extractionwith ethyl acetate, the organic phase was dried over sodium sulphate,filtered and evaporated to afford the title compound.

UPLC: 0.62 min, 256 [M+H]⁺

Intermediate 34 1,1-dimethylethyl{(1R)-1-[({6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinamine (Intermediate 33)was dissolved in DMF (0.5 mL) and added to a mixture of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (7.7 mg,0.038 mmol), DIPEA (0.008 mL, 0.047 mmol) and HATU (14.4 mg, 0.038 mmol)in 1 ml of DMF. The reaction mixture was then stirred at 50° C. for 2hours. After the removal of DMF, water was added and the mixture wasextracted with ethyl acetate. The gathered organic layers were driedover sodium sulphate, filtered and evaporated. The residue was thencharged on a silica gel column (Biotage SP1 system) and eluted withCyhexane/EtOAc (from all 10/0 to 7/3, then 7/3) to afford the titlecompound (4.4 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.24-8.17 (2H, m), 7.55-7.47 (1H, m),7.44-7.34 (1H, m), 7.08-7.00 (1H, m), 6.96-6.89 (1H, m), 5.00 (1H, br.s), 4.21-4.10 (1H, m), 2.94 (2H, q), 2.08-1.97 (1H, m), 1.80-1.70 (1H,m), 1.50 (9H, s), 1.38 (3H, t), 1.07 (3H, t). UPLC: 0.78 min, 441[M+H]+.

Intermediate 35(2R)-2-amino-N-{6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}butanamide

1,1-dimethylethyl{(1R)-1-[({6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 34, 4.4 mg) was dissolved in dichloromethane (1.0 ml) and,at 0° C., TFA (0.100 ml, 1.3 mmol) was added. The reaction mixture wasstirred at that temperature for 2 hours. After removal of the volatiles,the residue was charged on a SCX cartridge and eluted with DCM/MeOH/NH₃(2.0 M in MeOH) to afford the title compound.

UPLC: 0.53 min, 341 [M+H]+.

Intermediate 361-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-methyl-1-propanone

1,3-bis{[(methyloxy)methyl]oxy}benzene (Intermediate 35, 2.2 g) wasdissolved under nitrogen in THF (8.0 mL) and a solution of BuLi (8.32 mLof a 1.6 M solution in hexane, 13.3 mmol) was added. After stirring atroom temperature for 1 hour, cooling down to −78° C., 2-methylpropanoylchloride (4.65 ml, 44.4 mmol) was added and the reaction mixture wasstirred at that temperature for 1 hour. After quenching with water, thereaction mixture was extracted with ethyl acetate. The gathered organiclayers were dried over sodium sulphate, filtered and evaporated. Theresidue obtained was then charged on a silica gel column (Biotage SP1system) and eluted with Cyclohexane/EtOAc (from all 10/0 to 9/1, then9/1, then from 9/1 to 8/2, then 8/2) to afford the title compound (825mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.35-7.24 (1H, m), 6.87-6.77 (2H, m),5.18 (4H, s), 3.36 (6H, s), 3.01-2.90 (1H, m), 1.12-1.06 (6H, m).

The title compound was also prepared by the following alternative way:

1,3-bis{[(methyloxy)methyl]oxy}benzene (Intermediate 35, 1.099 g) wasdissolved under nitrogen in THF (5.0 mL) and a solution of BuLi (4.16 mLof a 1.6 M solution in hexane, 6.65 mmol) was added. After stirring atroom temperature for 1 hour, cooling down at −78° C., isobutyricanhydride (3.68 ml, 22.2 mmol) was added and the reaction mixture wasstirred at that temperature for 15 minutes. After quenching with water,the mixture was extracted with ethyl acetate. The gathered organiclayers were dried over sodium sulphate, filtered and evaporated. Theresidue obtained was then charged on a silica gel column and eluted withCyhexane/ethyl acetate (from all 10/0 to 9/1 then 9/1) to afford 759 mgof the title compound.

Intermediate 37 1-(2,6-dihydroxyphenyl)-2-methyl-1-propanone

1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-methyl-1-propanone(Intermediate 36, 1.58 g, 5.89 mmol) was dissolved in methanol (40.0 ml)and an aqueous solution of HCl (23.6 mL of a 2M aqueous solution, 47.1mmol) was added. The reaction mixture was refluxed for 1 hour. Afterquenching with water, the reaction mixture was extracted with ethylacetate. The gathered organic layers were dried over sodium sulphate,filtered and evaporated to afford the title compound (916 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 11.01 (2H, br. s), 7.15 (1H, t),6.39-6.34 (2H, m), 3.64-3.55 (1H, m), 1.09 (6H, d).

Intermediate 38 1-(2,6-dihydroxyphenyl)-2-methyl-1-propanone oxime

1-(2,6-dihydroxyphenyl)-2-methyl-1-propanone (Intermediate 37, 416 mg)was dissolved in pyridine (2.0 ml) and hydroxylamine hydrochloride (209mg, 3.0 mmol) was added. The reaction mixture was then stirred at roomtemperature overnight. After night, additional hydroxylaminehydrochloride (64 mg, 0.92 mmol) was added and the mixture was heated at100° C. for 3 hours. After removal of the volatiles, the crude wastreated with water and extracted with ethyl acetate. The gatheredorganic layers were dried over sodium sulphate, filtered and evaporatedto afford the title compound (370 mg).

UPLC: 0.47 min, 196 [M+H]+; 0.50 min, 196 [M+H]+.

Intermediate 39 1-(2,6-dihydroxyphenyl)-2-methyl-1-propanoneO-acetyloxime

1-(2,6-dihydroxyphenyl)-2-methyl-1-propanone oxime (Intermediate 38, 320mg) was stirred in acetic anhydride (0.928 ml, 9.84 mmol) at roomtemperature for 1 hour. After removal of the volatiles, the crude waswashed with water, filtered and dried. The residue obtained was chargedon a silica gel column (Biotage SP1 system) and eluted withCyhexane/ethyl acetate (from 100/0 to 0/100) to afford the titlecompound (127 mg).

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.42 (2H, br. s), 6.95 (1H, t), 6.31(2H, d), 2.91-2.77 (1H, m), 1.97 (3H, s), 1.11 (6H, d).

Intermediate 40 3-(1-methylethyl)-1,2-benzisoxazol-4-ol and2-(1-methylethyl)-1,3-benzoxazol-4-ol

1-(2,6-dihydroxyphenyl)-2-methyl-1-propanone O-acetyloxime (Intermediate39, 100 mg) was dissolved in pyridine (4.0 ml) and stirred at reflux for5 days. After the addition of an aqueous solution of HCl (5M), thereaction mixture was extracted 3 times with ethyl acetate and thegathered organic layers were washed with an aqueous solution of HCl(1M). The organic phase was then dried over sodium sulphate, filteredand evaporated. The residue obtained was charged on a silica gel column(Biotage SP1 system) and eluted with Cyhexane/EtOAc (from 1/0 to 1/1then 1/1, then from 1/1 to 0/1) to afford 13 mg of a mixture of thetitle compounds 3-(1-methylethyl)-1,2-benzisoxazol-4-ol and of2-(1-methylethyl)-1,3-benzoxazol-4-ol.

UPLC: 0.64 min, 178 [M+H]+; 0.65 min, 178 [M+H]+.

Intermediate 413-(1-methylethyl)-4-[(5-nitro-2-pyridinyl)oxy]-1,2-benzisoxazole

A mixture of 3-(1-methylethyl)-1,2-benzisoxazol-4-ol and2-(1-methylethyl)-1,3-benzoxazol-4-ol (Intermediate 40, 13 mg), wasdissolved in DMF (2.0 mL) and 2-chloro-5-nitropyridine (11.6 mg, 0.073mmol) was added, followed by potassium carbonate (30 mg, 0.22 mmol). Thereaction mixture was heated under microwave irradiation at 110° C. for 1hour. After the removal of the volatiles, the residue obtained wascharged on a silica gel column (Biotage SP1 system) and eluted withCyhexane/EtOAc (from 10/0 to 9/1, then 9/1, then from 9/1 to 8/2) toafford the title compound (8.7 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 9.08 (1H, d), 8.64-8.58 (1H, m),7.65-7.57 (1H, m), 7.55-7.48 (1H, m), 7.23-7.19 (1H, m), 7.08 (1H, d),3.24-3.13 (1H, m), 1.40 (6H, d). UPLC: 0.80 min, 300 [M+H]+.

Intermediate 426-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinamine

3-(1-methylethyl)-4-[(5-nitro-2-pyridinyl)oxy]-1,2-benzisoxazole(Intermediate 41, 8.7 mg) was dissolved in 3.0 ml of ethanol. Tin(II)chloride dihydrate (19.7 mg, 0.087 mmol) was added and the reactionmixture was stirred at 80° C. for 3 hours. After quenching with waterand extraction with ethyl acetate, the gathered organic layers weredried over sodium sulphate, filtered and evaporated to afford the titlecompound.

UPLC: 0.67 min, 270 [M+H]+.

Intermediate 431,1-dimethylethyl((1R)-1-{[(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinamine(Intermediate 42) was dissolved DMF (0.5 mL) and added to a mixture of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (7.09 mg,0.035 mmol), DIPEA (7.6 μL, 0.044 mmol) and HATU (13.26 mg, 0.035 mmol)in 1 ml of DMF. The reaction mixture was then stirred at 50° C. for 1hour. After removal of DMF, water was added and the mixture wasextracted with ethyl acetate. The gathered organic layers were driedover sodium sulpahte, filtered and evaporated. The residue was chargedon a silica gel column (Biotage SP1 system) and eluted withCyhexane/EtOAc (from 10/0 to 7/3, then 7/3) to afford the title compound(2 mg).

UPLC: 0.81 min, 455 [M+H]+.

Intermediate 44(2R)-2-amino-N-(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)butanamide

1,1-dimethylethyl((1R)-1-{[(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Intermediate 43, 2.0 mg) was dissolved in dichloromethane (1.0 mL) and,at 0° C., TFA (0.05 mL, 0.65 mmol) was added. The reaction mixture wasstirred at that temperature for 2 hours. After removal of the volatiles,the residue obtained was charged on a SCX cartridge and eluted withDCM/MeOH/NH3 (2.0 M in MeOH) to afford the title compound.

UPLC: 0.56 min, 355 [M+H]+.

Intermediate 45 1-(methyloxy)-3-{[(methyloxy)methyl]oxy}benzene

To a solution of 3-(methyloxy)phenol (10.38 g, 84 mmol) intetrahydrofurane (100 ml, SCRC) was added NaH (60% wt., 1.824 g, 76mmol, Aldrich) portionwise under ice-cooling. The reaction mixture wasstirred at room temperature for 1 hour and bromomethyl methyl ether (9.5g, 76 mmol, SCRC) was then added. The resulting mixture was stirred atroom temperature for 2 hours and water (50 ml) was added. The reactionmixture was extracted with ethyl acetate (2 times 50 ml, SCRC) and thecombined organic layers were dried over sodium sulphate, evaporated. Theresidue was purified by column chromatography on silica gel(EtOAc:PE=1:100) to afford the title compound (10.2 g) as a colorlessliquid.

Intermediate 46 2-iodo-1-(methyloxy)-3-{[(methyloxy)methyl]oxy}benzene

To a solution of 1-(methyloxy)-3-{[(methyloxy)methyl]oxy}benzene(Intermediate 45, 10 g, 59.5 mmol) in tetrahydrofurane (100 ml, SCRC)precooled to −78° C. was added dropwise BuLi (2.5 M in THF, 28.5 ml,71.3 mmol, SCRC), maintaining the inner temperature lower than −70° C.After the addition was complete, the mixture was stirred at −70° C. for2 hours and a solution of iodine (15.09 g, 59.5 mmol, SCRC) in THF (50ml, SCRC) was added dropwise. The resulting mixture was stirred for 2hours at room temperature and quenched with a saturated aqueous solutionof ammonium chloride (100 ml). The mixture was extracted with ethylacetate (3 times 300 ml, SCRC) and the combined organic layers weredried, evaporated and purified by silica gel chromatography with aseluents EtOAc:PE (1/100) to afford the title compound (16.2 g) as ayellow liquid.

Intermediate 47 2-iodo-3-(methyloxy)phenol

To a solution of 2-iodo-1-(methyloxy)-3-{[(methyloxy)methyl]oxy}benzene(Intermediate 46, 16.2 g, 55.1 mmol) in dichloromethane (100 ml, SCRC)was bubbled HCl (g) for 30 mins. TLC showed that the reaction wascompleted. The reaction mixture was poured into an aqueous saturatedsolution of NaHCO₃ (200 ml,) and extracted with dichloromethane (3×200ml, SCRC). The combined organic layers were dried, evaporated andpurified by column chromatography on silica gel (EtOAc:PE=1:50) toafford the title compound as a yellow liquid (10.3 g).

Intermediate 482-iodo-1-(methyloxy)-3-[(2-methyl-2-propen-1-yl)oxy]benzene

To a solution of 2-iodo-3-(methyloxy)phenol (Intermediate 47, 10.3 g) inDMF (100 ml, SCRC) was added NaH (60%, wt., 1.977 g, 49.4 mmol)portionwise. The reaction mixture was stirred at room temperature for 1hour and 3-chloro-2-methyl-1-propene (3.73 g, 41.2 mmol, Aldrich) wasadded. The resulting mixture was stirred at room temperature for 2 hoursand water (50 ml) was added. The reaction mixture was extracted withethyl acetate (3 times 200 ml, SCRC) and the combined organic layer weredried, evaporated and purified by silica gel chromatography with aseluents EtOAc/PE (1/30) to afford the title compound as a yellow liquid(11.6 g)

¹H-NMR (400 MHz, CDCl₃) δ ppm: 7.25 (1H, t), 6.52-6.47 (2H, m), 5.21(1H, s), 5.01 (1H, s), 4.49 (2H, s), 3.89 (3H, s), 1.87 (3H, s)

Intermediate 49 3,3-dimethyl-4-(methyloxy)-2,3-dihydro-1-benzofuran

To a solution of2-iodo-1-(methyloxy)-3-[(2-methyl-2-propen-1-yl)oxy]benzene(Intermediate 48, 6.08 g) in toluene (50 ml, SCRC) were added AIBN (3.61g, 21.99 mmol, SCRC) and tributylstannane (11.60 g, 40.0 mmol, Aldrich).The reaction mixture was heated at reflux for 3 hours and then cooled toroom temperature. Water (100 ml) was added and the mixture was extractedwith ethyl acetate (3 times 200 ml, SCRC). The combined organic layerswere dried, evaporated and purified by silica gel chromatography with aseluents EtOAc/PE (1/50) to afford the title compound as a yellow liquid(2.7 g).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.05 (1H, t), 6.50 (1H, d), 6.39 (1H,d), 4.14 (2H, s), 3.77 (3H, s), 1.34 (6H, s);

Intermediate 50 3,3-dimethyl-2,3-dihydro-1-benzofuran-4-ol

To a solution of 3,3-dimethyl-4-(methyloxy)-2,3-dihydro-1-benzofuran(Intermediate 49, 4.0 g) in dichloromethane (100 ml, SCRC) was addedBBr3 (6.37 ml, 67.3 mmol, SCRC) dropwise under ice-cooling. After theaddition was complete, the reaction mixture was stirred for 2 hours atroom temperature and then water (20 ml) was added. The resulting mixturewas extracted with ethyl acetate (3 times 100 ml, SCRC) and the combinedorganic layers were dried, evaporated and purified by silica gelchromatography with EtOAc/PE as eluents (1/20) to afford the titlecompound (2.8 g).

¹H-NMR (400 MHz, CDCl₃) δ ppm: 6.98-6.94 (1H, t), 6.41-6.39 (1H, dd),6.25-6.23 (1H, dd), 4.21 (2H, s), 1.45 (6H, s); MS_(—)2: 163 [M−H]−.

Intermediate 51 2-bromo-3-hydroxyphenyl acetate

To a solution of 2-bromo-1,3-benzenediol (3.028 g, 16.02 mmol) indichloromethane (70 ml), TEA (3.35 ml, 24.03 mmol) and acetic anhydride(1.512 ml, 16.02 mmol) were added under stirring. The reaction mixturewas stirred at room temperature overnight. The reaction was quenchedwith a saturated solution of ammonium chloride (100 ml), and extractedwith ethyl acetate (3 times 70 ml). The combined organic layers weredried over sodium sulphate, filtered and evaporated to afford the titlecompound as a black oil which was used directly used in the next step.(3.028 g)

UPLC_B: 0.41 min, 229 [M−H]−

Intermediate 52 2-bromo-3-[(2-methyl-2-propen-1-yl)oxy]phenyl acetate

To a solution of 2-bromo-3-hydroxyphenyl acetate (Intermediate 51, 3028mg) in acetonitrile (60 ml) potassium carbonate (3623 mg, 26.2 mmol) and3-bromo-2-methyl-1-propene (2123 mg, 15.73 mmol) were added. Thereaction mixture was stirred at room temperature overnight. The mixturewas washed with water (3 times 60 ml). The organic phase was separated,dried over sodium sulphate, filtered and evaporated. The residue waspurified by flash chromatography on silica gel using a 100 g-SNAP columnand cyclohexane/ethyl acetate from 100/0 to 80/20 as eluent to affordthe title compound as a colourless oil (2.324 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.27 (1H, t), 6.68 (1H, dd), 5.19 (1H,s), 5.04 (1H, s), 4.53 (2H, s), 2.38 (3H, s), 1.88 (3H, s); UPLC: 0.81min, 285 [M+H]+

Intermediate 53 3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl acetate

To a solution of 2-bromo-3-[(2-methyl-2-propen-b-yl)oxy]phenyl acetate(Intermediate 52, 2.324 g) in toluene (20 ml) AIBN (1.606 g, 9.78 mmol)and tributylstannane (4.73 g, 16.30 mmol) were added. The reactionmixture was stirred and heated at 100° C. for 2 hours, then was left atroom temperature for 4 hours. The reaction was quenched with water (60ml) and extracted with ethyl acetate (3 times 50 ml). The combinedorganic layers were dried over sodium sulphate, filtered and evaporated.The residue was purified by flash chromatography on silica gel using a100 g-SNAP column and cyclohexane/ethyl acetate from 100/0 to 70/30 aseluent to afford the title compound as a colourless oil (1.290 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.13 (1H, t), 6.68 (1H, d), 6.59 (1H, d),4.22 (2H, s), 2.33 (3H, s), 1.39 (6H, s). UPLC: 0.72 min, 207 [M+H]+

Intermediate 50 3,3-dimethyl-2,3-dihydro-1-benzofuran-4-ol

This is an alternative synthetic route to the one described previouslyfor Intermediate 50.

To a solution of 3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl acetate(Intermediate 53, 1.290 g) in methanol (50 ml) a solution of sodiumhydroxide (0.375 g, 9.38 mmol) in water (25.00 ml) was added. Thereaction mixture was stirred at room temperature for 30 minutes. Themixture was then acidified with HCl 5% until pH=5 and extracted withethyl acetate (3 times 50 ml). The combined organic layers were driedover sodium sulphate, filtered and evaporated. The residue was purifiedby flash chromatography on silica gel using a 25 g-SNAP column andcyclohexane/ethyl acetate from 100/0 to 80/20 as eluent to afford thetitle compound as a white solid (855 mg).

UPLC-MS: 0.65 min, 165 [M+H]+

Intermediate 543,3-dimethyl-4-[(4-nitrophenyl)oxy]-2,3-dihydro-1-benzofuran

To a solution of 3,3-dimethyl-2,3-dihydro-b-benzofuran-4-ol(Intermediate 50, 652 mg) and 1-fluoro-4-nitrobenzene (532 mg, 3.77mmol) in acetonitrile (30 ml, SCRC) was added potassium carbonate (552mg, 4 mmol, SCRC). The mixture was heated at reflux for 3 hours. Aftercooling, the reaction mixture was filtered and the filtrate wasevaporated to afford the title compound as a yellow liquid (0.95 g),which was directly used in the next step.

Intermediate 554-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]aniline

To a solution of3,3-dimethyl-4-[(4-nitrophenyl)oxy]-2,3-dihydro-1-benzofuran(Intermediate 54, 0.9 g) in tetrahydrofuran (20 ml, SCRC) and water (10ml) were added ammonium chloride (1.687 g, 31.5 mmol, SCRC) and zincpowder (1.031 g, 15.77 mmol, SCRC). The mixture was heated at 40° C. for2 hours and then filtered through a pad of celite. The filtrate waspartitioned with water (20 ml) and ethyl acetate (50 ml, SCRC). Theorganic layer was dried, evaporated and purified by columnchromatography on silica gel (EtOAc/PE=from 1/50 to 1/30) to afford thetitle compound as a yellow liquid (0.625 g, 76%).

MS_(—)2 (ESI): 256 [M+H]+

Intermediate 561,1-dimethylethyl[(1R)-2-({4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}amino)-1-methyl-2-oxoethyl]carbamate

To a solution of4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]aniline (Intermediate55, 255 mg) and N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (227 mg,1.199 mmol, SCRC) in DMF (10 ml, SCRC) were added HATU (570 mg, 1.498mmol, SCRC) and DIPEA (0.523 ml, 3.00 mmol, SCRC). The mixture washeated at 100° C. in microwave for 1 hour. Water (20 ml) was added tothe mixture and it was extracted with ethyl acetate (3 times 50 ml,SCRC). The combined organic layers were dried, evaporated and purifiedby column chromatography on silica gel (EtOAc/PE=from 1/50 to 1/20) toafford the title compound as a yellow solid (328 mg).

MS_(—)2 (ESI): 371 [M−55]+

Intermediate 57

N¹-{4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}-D-alaninamide

To a solution of1,1-dimethylethyl[(1R)-2-({4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 56, 325 mg) in ethyl acetate (20 ml, SCRC) was bubbledinto HCl (gas) for 0.5 hours. The reaction mixture was neutralized witha saturated aqueous solution of sodium carbonate pH=7 and it wasextracted with ethyl acetate (3 times 100 ml, SCRC). The combinedorganic layers were dried and evaporated to afford the title compound asa yellow liquid (215 mg).

MS_(—)2 (ESI): 327 [M−H]+

Intermediate 582-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-nitropyridine

In a large microwave vial, 2-chloro-5-nitropyridine (386 mg, 2.436 mmol)was dissolved in 4 ml of dimethylformamide.3,3-dimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 57, 400 mg) andpotassium carbonate (2.02 g, 14.62 mmol) were added. The reactionmixture was heated under microwave irradiation during 30 minutes at 110C (Biotage Initiator). The reaction mixture was filtered. The filtratedsolid was washed with dichloromethane (30 ml). The volatiles wereevaporated under vacuum. The residue was purified by silica gelchromatography (Companion instrument, 120 g cartridge) with cyclohexaneas eluent to afford the title compound (470 mg).

¹H NMR (400 MHz, MeOD): δ ppm 9.00 (1H, d), 8.59 (1H, dd), 6.99-7.33(2H, m), 6.66 (1H, d), 6.57 (1H, d), 4.20 (2H, s), 1.17-1.35 (6H, m);UPLC: 0.88 min, 287 [M+H]+

Intermediate 596-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine

2-{[3-(1-methylethyl)phenyl]oxy}-5-nitropyridine (Intermediate 58, 465mg) was dissolved in ethanol (8 ml). Hydrazine monohydrate (156 mg, 3.25mmol, 2 equiv) and palladium on carbon (121 mg, 0.114 mmol) were added.The reaction mixture was heated at reflux under argon during 3 hours.The reaction was cooled down and then filtered on celite. The organicphase was evaporated under vacuum. Evaporation afforded the titlecompound as a yellow oil (300 mg).

¹H NMR (400 MHz, MeOD): δ ppm 7.65 (1H, d), 7.21 (1H, dd), 7.03 (1H, t),6.73 (1H, d), 6.51 (1H, d), 6.26 (1H, d), 4.21 (2H, s), 1.40 (6H, s);UPLC: 0.64 min, 257 [M+H]+

Intermediate 601,1-dimethylethyl[(1R)-2-({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alanine (35.4mg, 0.156 mmol) in dry N,N-dimethylformamide (3 ml), DIPEA (0.041 ml,0.264 mmol) and then HATU (71 mg, 0.187 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature underargon. Then6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 59, 40 mg) was added and the reaction mixture was stirred3 hours at 60° C. under argon. The reaction mixture was evaporated. Theresidue obtained was purified by silica gel chromatography (Companionsystem, 2×12 g=24 g cartridge) with a gradient cyclohexane/ethyl acetatefrom 100/0 to 70/30. This afforded the title compound (43 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.35 (1H, d), 8.08 (1H, dd), 7.11 (1H, t),6.93 (1H, d), 6.60 (1H, d), 6.45 (1H, d), 4.21 (2H, s), 4.15-4.04 (1H,m), 1.47-1.33 (18H, m); UPLC: 0.78 min, 428 [M+H]+

Intermediate 61

N¹-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-D-alaninamide

To a solution of1,1-dimethylethyl[(1R)-2-({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamate(Intermediate 60, 35 mg) in dry dichloromethane (3 ml), TFA (0.189 ml,2.456 mmol) was slowly added at 0° C. and the reaction mixture wasstirred for 1.5 hours at room temperature. The solvent and the excess ofTFA were evaporated and the residue was purified with an SCX cartridge.The cartridge was washed with 3 CV of methanol, then the compound wasadsorbed on the cartridge, washed with 5 CV of methanol and desorbedwith 2 CV of methanolic ammonia (1N). This afforded the title compound(32 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.41 (1H, d), 8.14 (1H, dd), 7.16 (1H, t),6.97 (1H, d), 6.64 (1H, d), 6.48 (1H, d), 4.24 (2H, s), 3.62 (1H, m),1.50-1.38 (9H, m); UPLC: 0.55 min, 328 [M+H]+

Intermediate 62 1,1-dimethylethyl{(1R)-1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

The title compound was made in a similar fashion to the preparation ofIntermediate 60 replacing N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-alaninewith (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (66.6mg). This afforded 78 mg of the title compound.

¹H NMR (400 MHz, MeOD): δ ppm 8.37-8.32 (1H, m), 8.07 (1H, dd), 7.11(1H, t), 6.93 (1H, d), 6.60 (1H, d), 6.44 (1H, d), 4.21 (2H, s),4.12-4.05 (1H, m), 1.91-1.78 (1H, m), 1.74-1.64 (1H, m), 1.50-1.39 (12H,m), 1.37-1.30 (6H, s); UPLC: 0.81 min, 442 [M+H]+

Intermediate 63(2R)-2-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide

The title compound was made in a similar fashion to the preparation ofIntermediate 61 replacing1,1-dimethylethyl[(1R)-2-({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1-methyl-2-oxoethyl]carbamatewith 1,1-dimethylethyl{(1R)-1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

(Intermediate 62, 74 mg). This afforded 60 mg of the title compound.

¹H NMR (400 MHz, MeOD): δ ppm 8.38 (1H, d), 8.12 (1H, dd), 7.11 (1H, t),6.95 (1H, d), 6.61 (1H, d), 6.46 (1H, d), 4.21 (2H, s), 3.41 (1H, m),1.81 (1H, m), 1.70 (1H, m), 1.35 (6H, s), 1.00 (3H, t); UPLC: 0.55 min,342 [M+H]+

Intermediate 642-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-nitropyrimidine

To a solution of 3,3-dimethyl-2,3-dihydro-1-benzofuran-4-ol(Intermediate 50, 724 mg) in dry N,N-Dimethylformamide (40 mL) potassiumcarbonate and 2-chloro-5-nitropyrimidine (774 mg, 4.85 mmol) were added.The reaction mixture was stirred at room temperature overnight. Thereaction was quenched with water (40 ml) and extracted with ethylacetate (3×40 ml). The combined organic layers were washed with brine(2×50 ml), separated, dried over sodium sulphate, filtered andevaporated. The residue was purified by flash chromatography on silicagel using a column SNAP 50 g and cyclohexane/ethyl acetate as eluentsfrom 100/0 to 70/30 to afford the title compound (1.257 g) as a yellowoil.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.46 (2H, s), 7.22 (1H, t), 6.73 (2H,dd), 4.23 (2H, s), 1.24 (6H, s); UPLC: 0.75 min, 288 [M+H]+

Intermediate 652-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine

To a solution of2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-nitropyrimidine(Intermediate 64, 1.257 g) in a mixture tetrahydrofuran/water (30ml/15.00 ml) iron (1.222 g, 21.88 mmol) and ammonium chloride (1.170 g,21.88 mmol) were added. The reaction mixture was stirred at roomtemperature for 48 hours. The catalyst was filtered off, and thefiltrate was extracted with ethyl acetate (3×50 ml). The combinedorganic layers were dried over sodium sulphate, filtered and evaporated.The residue was recrystallized from ethyl acetate to afford the titlecompound (768 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.97 (2H, s), 6.94-7.16 (1H, m), 6.58(1H, d), 6.44 (1H, d), 4.18 (2H, s), 3.32 (2H, br. s.), 1.25 (6H, s);UPLC: 0.60 min, 258 [M+H]+

Intermediate 66 1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (20.14 mg,0.099 mmol) in N,N-dimethylformamide (1.5 mL) DIPEA (0.029 mL, 0.165mmol) and TBTU (33.9 mg, 0.106 mmol) were added. The mixture reactionwas stirred during 15 minutes at room temperature, then2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine(Intermediate 65, 17 mg) was added. The reaction mixture was stirredduring 48 hours at room temperature. The mixture was diluted with ethylacetate (5 ml) and washed with brine (3×5 ml). The organic layer wasseparated, dried over sodium sulphate, filtered and evaporated. Theresidue was purified by silica gel flash chromatography using a 10g-SNAP column and cyclohexane/ethyl acetate from 100/0 to 40/60 aseluent to afford the title compound as a white solid (4.4 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.78 (2H, s), 7.17 (1H, t), 6.71 (1H, d),6.61 (1H, d), 4.97 (1H, d), 4.24 (2H, s), 4.18-4.11 (1H, m), 2.05-1.97(1H, m), 1.77-1.70 (1H, m), 1.49 (9H, s), 1.37 (6H, s), 1.05 (3H, t).UPLC-MS: 0.78 min, 443 [M+H]+

Intermediate 67(2R)-2-amino-N-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}butanamide

To a solution of 1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate(Intermediate 66, 4.4 mg) in dichloromethane (1 ml) cooled to 0° C. TFA(0.019 ml, 0.249 mmol) was added dropwise. The mixture reaction wasstirred at 0° C. for 1.5 hours. The solvent and the TFA were evaporated.The mixture was diluted with dichloromethane (5 ml) and neutralized withan aqueous saturated solution of NaHCO₃ (5 ml). The organic layer wasseparated, dried over sodium sulphate, filtered end evaporated to affordthe title compound (3 mg) which was directly used in the next step

UPLC-MS: 0.98 min, 343 [M+H]+

Intermediate 681-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acid

To a solution of 1-aminocyclobutanecarboxylic acid (626 mg, 5.44 mmol)in 5.6 ml of 1 M aqueous sodium hydroxide and 4 ml of methanol was addedBoc-anhydride (1.425 g, 6.53 mmol) at 0° C. The reaction mixture waswarmed to room temperature and stirred for 12 hours. After most of themethanol was evaporated, the solution was acidified to pH 2 with 1 M HCland extracted with ethyl acetate. The organic extracts were combined andwashed with brine. Evaporation of the solvent afforded the titlecompound (1.09 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.21 (1H, s), 7.44 (1H, s), 2.29-2.47(2H, m), 2.09 (2H, q), 1.74-1.94 (2H, m), 1.36 (9H, s); UPLC: 0.56 min,216 [M+H]+

Intermediate 69 1,1-dimethylethyl{1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]cyclobutyl}carbamate

To a solution of1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acid(Intermediate 68, 20.16 mg) in dry N,Ndimethylformamide (3 ml), DIPEA(20.44 μl, 0.117 mmol) and then HATU (35.6 mg, 0.094 mmol) were addedand the reaction mixture was stirred for 15 minutes at room temperatureunder argon. Then6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamineIntermediate 59, 20 mg) was added and the reaction mixture was stirredat 60° C. under argon during 12 hours. The reaction mixture was cooleddown and a prestirred (15 min) solution of HATU (1 equiv),1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acid (1equiv) and DIPEA (1 equiv) in 1 mL of dry DMF was added. The reactionmixture was heated under argon an additional 12 hours at 60° C. Thereaction mixture was evaporated. The residue obtained was purified onsilica gel (Companion instrument) with cyclohexane/ethylacetate aseluents from 100/0 to 70/30. This afforded the title compound (14 mg)

¹H NMR (400 MHz, MeOD): δ ppm 8.29 (1H, s), 8.13-7.86 (1H, m), 7.11 (1H,t), 7.04-6.76 (1H, m), 6.60 (1H, d), 6.45 (1H, d), 4.21 (2H, s)2.85-2.52 (2H, m), 2.26-2.09 (2H, m), 2.07-1.80 (2H, m), 1.45 (6H, s),1.35 (9H, s); UPLC: 0.95 min, 454 [M+H]+

Intermediate 701-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}cyclobutanecarboxamide

To a solution of 1,1-dimethylethyl{1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]cyclobutyl}carbamate(Intermediate 69, 23.5 mg) in dry dichloromethane (3.5 ml), TFA (159 μl,2.07 mmol) was slowly added at 0° C. and the reaction mixture wasstirred for 2 hours at room temperature. The solvent and the excess ofTFA were evaporated and the residue was purified with an SCX cartridge.The cartridge was washed with 3 CV of methanol, then the compound wasadsorbed on the cartridge, washed with 5 CV of methanol and desorbedwith 2 CV of methanolic ammonia (1N). This afforded the title compound(18 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.43 (1H, d), 8.11 (1H, dd), 7.11 (1H, t),6.94 (1H, d), 6.61 (1H, d), 6.45 (1H, d), 4.20 (2H, s), 2.75-2.70 (2H,m), 2.18-2.00 (4H, m), 1.34 (6H, m); UPLC: 0.82 min, 354 [M+H]+

Intermediate 711-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclopropanecarboxylic acid

The title compound (998 mg) was made in a similar fashion to thepreparation of Intermediate 68 replacing 1-aminocyclobutanecarboxylicacid WITH 1-aminocyclopropanecarboxylic acid (550 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.26 (1H, s), 7.40 (1H, s), 1.38 (9H,s), 1.26 (2H, m), 0.96 (2H, m); UPLC: 0.52 min, 202 [M+H]+

Intermediate 72 1,1-dimethylethyl{1-≡({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]cyclopropyl}carbamate

The title compound (14 mg) was made in a similar fashion to thepreparation of Intermediate 69 replacing1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclobutanecarboxylic acidwith 1-({[(1,1-dimethylethyl)oxy]carbonyl}amino)cyclopropanecarboxylicacid (Intermediate 71, 18.84 mg, 0.094 mmol).

¹H NMR (400 MHz, MeOD): δ ppm 8.30 (1H, s), 8.08-7.93 (1H, m), 7.11 (1H,t) 6.92 (1H, d), 6.60 (1H, d), 6.45 (1H, d), 4.22 (2H, s), 1.58-1.51(2H, m), 1.45 (9H, s), 1.35 (6H, s), 1.12 (2H, m); UPLC: 0.78 min, 440[M+H]+

Intermediate 731-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}cyclopropanecarboxamide

The title compound (10 mg) was made in a similar fashion to thepreparation of Intermediate 70 replacing 1,1-dimethylethyl{1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]cyclobutyl}carbamatewith1,1-dimethylethyl{1-[({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]cyclopropyl}carbamate(Intermediate 72, 13 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.39 (1H, d), 8.07 (1H, dd), 7.10 (1H, t),6.92 (1H, d), 6.59 (1H, d), 6.44 (1H, d), 4.20 (2H, s), 1.40-1.31 (8H,m), 1.04-0.96 (2H, m); UPLC: 0.51 min, 340 [M+H]+

Intermediate 741,1-dimethylethyl[2-({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

To a solution of (N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(39.0 mg, 0.192 mmol) in dry N,Ndimethylformamide (3 mL), DIPEA (0.042mL, 0.240 mmol) and then HATU (73.0 mg, 0.192 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature underargon. Then6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 59, 41 mg) was added and the reaction mixture was stirredat 60° C. under argon. The reaction was left under heating 4 hours andwas stop before completion. The reaction mixture was evaporated. Theresidue obtained was purified on silica gel (Companion instrument) witha gradient cyclohexane/ethylacetate from 100/0 to 70/30. This affordedthe title compound (13 mg).

¹H NMR (400 MHz, MeOH): δ ppm 9.66 (1H, s), 8.28 (1H, s), 8.04 (1H, s),7.80-7.51 (1H, m), 7.11 (1H, t), 6.92 (1H, d), 6.60 (1H, d), 6.45 (1H,d), 4.21 (2H, s), 1.48 (6H, s), 1.43 (6H, s), 1.35 (9H, s); UPLC: 0.79min, 442 [M+H]+

Intermediate 75

N¹-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-2-methylalaninamide

To a solution of1,1-dimethylethyl[2-({6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 74, 11 mg) in dry dichloromethane (2 ml), TFA (0.077 ml,0.997 mmol) was slowly added at 0° C. and the reaction mixture wasstirred for 2 hours at room temperature. The solvent and the excess ofTFA were evaporated and the residue was purified with an SCX cartridge.The cartridge was washed with 3 CV of methanol and then the compound wasadsorbed on the cartridge, washed with 5 CV of methanol and desorbedwith 2 CV of methanolic ammonia (1N). This afforded the title compound(8.5 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.39 (1H, d), 8.07 (1H, dd), 7.11 (1H, t),6.92 (1H, d), 6.59 (1H, d), 6.43 (1H, d), 4.21 (2H, s), 1.45 (6H, s),1.35 (6H, s); UPLC: 0.50 min, 342 [M+H]+

Intermediate 76 N-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-D-valine

To a solution of 3-methyl-D-valine (900 mg, 6.86 mmol) in 7 ml of 1 Maqueous sodium hydroxide and 7 ml of methanol was added Boc-anhydride(1.797 g, 8.23 mmol) at 0° C. The reaction mixture was warmed to roomtemperature and stirred overnight. After most of the methanol wasevaporated, the solution was acidified to pH 2 with an aqueous solutionof HCl (1M) and extracted 3 times with ethylacetate (3×20 ml). Theorganic layers were combined and washed with brine (2×5 ml). Evaporationof the solvent afforded the title compound as a white solid (1.36 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.44 (1H, s), 6.82 (1H, d), 3.76 (1H,d), 1.38 (9H, s), 0.93 (9H, s); UPLC: 0.64 min, 232 [M+H]+

Intermediate 77 1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]-2,2-dimethylpropyl}carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-3-methyl-D-valine(Intermediate 76, 53.9 mg) in dry N,Ndimethylformamide (1 ml), DIPEA(50.9 μl, 0.292 mmol) and then HATU (102 mg, 0.268 mmol) were added andthe reaction mixture was stirred for 15 minutes at room temperatureunder argon. Then2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine(Intermediate 65, 30 mg) was added and the reaction mixture was stirredat 60° C. under argon during 12 hours. The reaction was quenched withbrine (1 ml), diluted with water (2 ml) and extracted with ethyl acetate(2×5 ml). The organic layer was dried over sodium sulphate andevaporated. The residue obtained was purified on silica gel (Companioninstrument) with a gradient cyclohexane/ethylacetate 100/0 to 70/30.This afforded the title compound (17 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.74 (2H, s), 7.14 (1H, t), 6.67 (1H, d),6.57 (1H, d), 4.53 (1H, d), 4.20 (2H, s), 1.40 (6H, s) 1.06 (9H, s),0.98 (9H, s); UPLC: 0.83 min, 371 [M+H]+

Intermediate 78N¹-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-3-methyl-D-valinamide

To a solution of 1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]-2,2-dimethylpropyl}carbamate(Intermediate 77, 13 mg) in dry dichloromethane (0.5 ml) cooled to 0°C., TFA (85 μl, 1.105 mmol) was added dropwise and the solution wasstirred for 3 hours at that temperature. The volatiles were evaporated.The residue was dissolved with dichloromethane (2 ml) and an aqueoussaturated solution of NaHCO₃ was added (4 ml). The layers were separatedand the aqueous layer was extracted twice with dichloromethane. Thegathered organic layers were dried over sodium sulphate and evaporatedto afford the title compound (10.9 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.68 (2H, s), 7.14 (1H, t), 6.68 (1H, d),6.59 (1H, d), 4.21 (2H, s), 3.30 (1H, s), 1.70 (2H, br s), 1.35 (6H, s)1.07 (9H, s); UPLC: 0.53 min, 377 [M+H]+

Intermediate 79 1,3-bis{[(methyloxy)methyl]oxy}benzene

To a solution of 1,3-benzenediol (1.5 g, 13.62 mmol) in dryN,N-Dimethylformamide (13.62 ml) at 0° C. sodium hydride (0.981 g, 40.9mmol) was added and the reaction mixture was stirred for 15 minutes atthe same temperature. MOM-Cl (3.10 ml, 40.9 mmol) was quickly added andthe reaction mixture was stirred for 1 hour while the temperature wasallowed to reach room temperature. The reaction was quenched with brine(20 ml) and extracted with ethyl acetate (3×50 ml). The organic layerwas washed with brine (2×30 ml), dried over sodium sulphate, filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 50 g SNAP column and cyclohexaneto cyclohexane/ethyl acetate 8:2 as eluents affording the title compound(1.59 g, 8.02 mmol) as a colourless oil.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.16-7.23 (1H, d), 6.69-6.64 (3H, m),5.17 (4H, s), 3.38 (6H, s).

Intermediate 80 ethyl(2,6-bis{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate

To a solution of 1,3-bis{[(methyloxy)methyl]oxy}benzene (Intermediate79, 2.19 g) in dry tetrahydrofuran (10 ml) at room temperature BuLi 1.6Min hexane (8.29 ml, 13.26 mmol) was added and the reaction mixture wasstirred for 30 minutes at the same temperature. The mixture was cooledto −78° C. and it was added (via cannulation) to a solution of ethylchloro(oxo)acetate (2.263 g, 16.57 mmol) in dry tetrahydrofuran (10 ml)at −78° C. The reaction mixture was stirred at −78° C. for 30 minutes.The reaction was quenched with an aqueous saturated solution of ammoniumchloride (10 ml) and extracted with ethyl acetate (2×30 ml). Combinedorganic layers were dried over sodium sulphate, filtered and evaporated.The residue was purified by flash chromatography (Biotage system) onsilica gel using a 100 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 8:2 as eluent affording the title compound asa light yellow oil (1.75 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.46 (1H, t), 6.87 (2H, d), 5.20 (4H,s), 4.29 (2H, q), 3.34 (6H, s), 1.27 (3H, t).

Intermediate 81 ethyl2-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-propenoate

To a suspension of methyltriphenylphosphonium bromide (3.13 g, 8.75mmol) in dry tetrahydrofuran (30 ml) at 0° C. KHMDS (1.745 g, 8.75 mmol)was slowly added and the reaction mixture was stirred for 15 minutes at0° C. and for 45 minutes at room temperature. The reaction mixture wascooled to 0° C. and a solution of ethyl(2,6-bis{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate (Intermediate 80,1.74 g) in dry tetrahydrofuran (10 mL) was slowly added and the reactionmixture was stirred for 2 hours at 0° C. The reaction was quenched withan aqueous saturated solution of ammonium chloride (10 ml), diluted withwater (20 ml) and extracted with ethyl acetate (2×50 ml). The organiclayer was dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography (Biotage system) on silicagel using a 100 g SNAP column and cyclohexane to cyclohexane/ethylacetate 8:2 as eluents affording the title compound as a colourless oil(1.37 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.21 (1H, t), 6.78 (2H, d), 6.44 (1H,d), 5.74 (1H, d), 5.12 (4H, s), 4.12 (2H, q), 3.32 (6H, s), 1.17 (3H,t).

Intermediate 82 ethyl1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate

To a solution of trimethylsulfoxonium iodide (1.805 g, 8.20 mmol) in drydimethyl sulfoxide (20 mL) sodium hydride 60% dispersion in mineral oil(0.310 g, 7.75 mmol) was added and the reaction mixture was stirred for1 hour at room temperature. A solution of ethyl2-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)-2-propenoate (Intermediate 81,1.35 g) in dry dimethyl sulfoxide (10 mL) was slowly added and thereaction mixture was stirred for 1 hour at room temperature. Thereaction was quenched with an aqueous saturated solution of ammoniumchloride (10 ml), diluted with water (20 ml) and extracted with ethylacetate (2×50 ml). The organic layer was washed with water (50 ml),dried over sodium sulphate, filtered and evaporated. The residue waspurified by flash chromatography (Biotage system) on silica gel using a50 g SNAP column and cyclohexane to cyclohexane/ethyl acetate 8:2 aseluents affording the title as a colourless oil (1.14 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.15 (1H, t), 6.71 (2H, d), 5.18 (4H,s), 3.97 (2H, q), 3.36 (6H, s), 1.53-1.58 (2H, m), 1.09-1.14 (2H, m),1.04 (3H, t).

Intermediate 832-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol

To a solution of ethyl1-(2,6-bis{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate(Intermediate 82, 490 mg) in ethanol (10 ml) HCl 2N in water (0.789 mL,1.579 mmol) was added and the reaction mixture was stirred overnight at50° C. Toluene (20 mL) was added and the combined solvents were removedunder reduced pressure. The residue was re-suspended in toluene (20 ml)and the solvent evaporated. The obtained residue was dissolved in drytetrahydrofuran (20 ml), the mixture was cooled to 0° C. and NaH 60%dispersion in mineral oil (126 mg, 3.16 mmol) was added and the reactionmixture was stirred for 30 minutes at the same temperature. MOM-Cl(0.120 mL, 1.579 mmol) was then added and the reaction mixture wasstirred for 2 hours at 0° C. LiAlH4 (1M in THF, 1.579 ml, 1.579 mmol)was added and the reaction mixture was further stirred for 1 hour at thesame temperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (10 ml), diluted with water (10 ml) andextracted with ethyl acetate (2×50 ml). Combined organic layers weredried over sodium sulphate, filtered and evaporated and the residue waspurified by flash chromatography (Biotage system) on silica gel using a25 g SNAP column and cyclohexane to cyclohexane/ethyl acetate 7:3 aseluents affording the title compound as a colourless oil (191 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.90 (1H, br.s) 6.96 (1H, t), 6.50 (1H,d), 6.45 (1H, d), 5.16 (2H, s), 4.93 (1H, br.s), 3.45 (2H, s), 3.40 (3H,s), 0.86-0.93 (2H, m), 0.56-0.62 (2H, m); UPLC: 0.59 min, 225 [M+H]+.

Intermediate 844-{[(methyloxy)methyl]oxy}spiro[1-benzofuran-3,1′-cyclopropane]

To a solution of2-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol(Intermediate 83, 190 mg) in dry tetrahydrofuran (10 ml)triphenylphosphine (333 mg, 1.271 mmol) was added and the reactionmixture was stirred until complete dissolution of PPh3. DIAD (0.198 ml,1.017 mmol) was then added dropwise and the reaction mixture was stirredfor 30 minutes at room temperature The solvent was removed under reducedpressure. The residue was purified by flash chromatography (Biotagesystem) on silica gel using a 25 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 9:1 as eluents affording the title compound asa light yellow oil (120 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 6.97 (1H, t), 6.51 (1H, d), 6.43 (1H,d), 5.12 (2H, s), 4.40 (2H, s), 3.35 (3H, s), 1.43-1.48 (2H, m),0.85-0.90 (2H, m); UPLC_B: 0.88 min, 207 [M+H]+.

Intermediate 85 spiro[1-benzofuran-3,1′-cyclopropan]-4-ol

To a solution of4-{[(methyloxy)methyl]oxy}spiro[1-benzofuran-3,1′-cyclopropane](Intermediate 84, 118 mg) in methanol (5 ml), HCl 2N in water (0.286 mL,0.572 mmol) was added and the reaction mixture was stirred overnight at50° C. Combined solvents were removed under reduced pressure and theresidue was re-dissolved in toluen (10 ml) and the solvent was removed.The residue was purified by flash chromatography (Biotage system) onsilica gel using a 10 g SNAP column and cyclohexane to cyclohexane/ethylacetate 7:3 as eluents affording the title compound as a white solid (70mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.28 (1H, s), 6.81 (1H, t), 6.24 (1H,d), 6.22 (1H, d), 4.34 (2H, s), 1.40-1.45 (2H, m), 0.77-0.82 (2H, m).

Intermediate 865-nitro-2-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)pyridine

To a solution of spiro[1-benzofuran-3,1′-cyclopropan]-4-ol (Intermediate85, 70 mg) in dry N,N-dimethylformamide (2 ml) potassium carbonate (89mg, 0.647 mmol) and then 2-chloro-5-nitropyridine (75 mg, 0.475 mmol)were added and the reaction mixture was stirred for 3 hours at 100° C.The reaction was quenched with brine (1 ml), diluted with water (2 ml)and extracted with ethyl acetate (3×10 ml). The organic layer was driedover sodium sulphate, filtered and evaporated. The residue was purifiedby flash chromatography (Biotage system) on silica gel using a 10 g SNAPcolumn and cyclohexane to cyclohexane/ethyl acetate 9:1 as eluentsaffording the title compound as a white solid (100 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.05 (1H, d), 8.63 (1H, dd), 7.23 (1H,d), 7.13 (1H, t), 6.73 (1H, d), 6.60 (1H, d), 4.45 (2H, s), 1.05-1.10(2H, m), 0.88-0.93 (2H, m); UPLC: 0.79 min, 285 [M+H]+.

Intermediate 876-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine

To a solution of5-nitro-2-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)pyridine(Intermediate 86, 99 mg) in tetrahydrofuran (5 ml)/water (2.5 ml) iron(97 mg, 1.741 mmol) and then ammonium chloride (93 mg, 1.741 mmol) wereadded and the reaction mixture was stirred for 4 hours at roomtemperature. The catalyst was filtered off and the residue was dilutedwith an aqueous saturated solution of NaHCO3 (5 ml) and extracted withethyl acetate (3×10 ml). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 1:1 aseluents affording the title compound as a light yellow solid (85 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.52 (1H, d), 7.06 (1H, dd), 6.97 (1H,t), 6.70 (1H, d), 6.53 (1H, d), 6.23 (1H, d), 5.08 (2H, s), 4.43 (2H,s), 1.28-1.33 (2H, m), 0.86-0.91 (2H, m); UPLC: 0.62 min, 255 [M+H]+.

Intermediate 881,1-dimethylethyl[(1R)-1-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (94 mg,0.462 mmol) in dry N,N-Dimethylformamide (2 mL) DIPEA (0.115 mL, 0.661mmol) and then TBTU (159 mg, 0.496 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine(Intermediate 87, 84 mg) was added and the reaction mixture was stirredfor 6 hours at the same temperature. The reaction was quenched withbrine (2 ml), diluted with water (5 ml) and extracted with ethyl acetate(2×10 ml). The organic layer was washed with ice cold brine (2×5 ml),dried over sodium sulphate, filtered and evaporated. The residue waspurified by flash chromatography (Biotage system) on silica gel using a10 g SNAP column and cyclohexane to cyclohexane/ethyl acetate 7:3 aseluents affording the title compound as a colourless oil (130 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 10.14 (1H, br.s), 8.32 (1H, d), 8.08(1H, dd), 7.02-7.09 (2H, m), 6.96 (1H, d), 6.63 (1H, d), 6.42 (1H, d),4.44 (2H, s), 3.93-4.01 (1H, m), 1.52-1.75 (2H, m), 1.39 (9H, s),1.15-1.22 (2H, m), 0.85-0.95 (5H, m); UPLC: 0.80 min, 440 [M+H]+.

Intermediate 89(2R)-2-amino-N-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]butanamide

To a solution of1,1-dimethylethyl[(1R)-1-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate(Intermediate 88, 128 mg) in dry dichloromethane (3 ml) at 0° C. TFA(0.9 mL, 11.68 mmol) was slowly added and the reaction mixture wasstirred for 2 hours at the same temperature. The reaction was dilutedwith dichloromethane (10 ml) and an aqueous saturated solution of NaHCO₃was added while the pH was allowed to reach ˜8. Two phases wereseparated and the aqueous layer was re-extracted with dichloromethane(10 ml). The organic layers were combined, dried over sodium sulphate,filtered and evaporated affording the title compound as a colourless oil(92 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.37 (1H, d), 8.13 (1H, dd), 7.05 (1H,t), 6.95 (1H, d), 6.63 (1H, d), 6.42 (1H, d), 4.44 (2H, s), 3.24 (1H,m), 1.61-1.72 (1H, m), 1.44-1.55 (1H, m), 1.16-1.21 (2H, m), 0.91 (3H,t), 0.86-0.91 (2H, m); UPLC_B: 0.74 min, 340 [M+H]+.

Intermediate 90 1,1-dimethylethyl(1,1-dimethyl-2-oxo-2-{[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}ethyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(80 mg, 0.393 mmol) in dry N,N-dimethylformamide (1.5 mL) DIPEA (0.096mL, 0.551 mmol) and then HATU (150 mg, 0.393 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature Thissolution was added to a solution of6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine(Intermediate 89, 40 mg) in dry N,N-dimethylformamide (0.5 ml) and thereaction mixture was stirred overnight at room temperature. The reactionwas quenched with water (2 ml), diluted with brine (10 ml) and extractedwith ethyl acetate (2×20 ml). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 1:1 aseluents affording the title compound as a white solid (52 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.62 (1H, br.s), 8.24-8.42 (1H, br.m),8.05 (1H, d), 6.98-7.10 (2H, m), 6.92 (1H, d), 6.61 (1H, d), 6.40 (1H,d), 4.44 (2H, s), 1.42 (6H, s), 1.36 (9H, s), 1.15-1.21 (2H, m),0.85-0.91 (2H, m); UPLC: 0.81 min, 440 [M+H]+.

Intermediate 912-methyl-N¹-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]alaninamide

To a solution of 1,1-dimethylethyl(1,1-dimethyl-2-oxo-2-{[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}ethyl)carbamate(Intermediate 90, 50 mg) in dry dichloromethane (4 mL) at 0° C. TFA (1ml, 12.98 mmol) was slowly added and the reaction mixture was stirredfor 2 hours at the same temperature. The reaction was diluted withdichloromethane (10 ml) and an aqueous saturated solution of NaHCO₃ wasadded while the pH was allowed to reach ˜8. Two phases were separatedand the organic layer was dried over sodium sulphate, filtered andevaporated affording the title compound (35 mg) as a colourless oil.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.40 (1H, d), 8.15 (1H, dd), 7.04 (1H,t), 6.94 (1H, d), 6.62 (1H, d), 6.41 (1H, d), 4.43 (2H, s), 1.28 (6H,s), 1.15-1.20 (2H, m), 0.86-0.91 (2H, m); UPLC: 0.56 min, 340 [M+H]+.

Intermediate 921,1-dimethylethyl[(1R)-1-methyl-1-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate

6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinamine(Intermediate 91, 127 mg),N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline (Nagase & Co Ltd, 109mg, 0.499 mmol), DIPEA (0.131 mL, 0.749 mmol) and HATU (247 mg, 0.649mmol) were dissolved in dry N,N-dimethylformamide (3 ml) and the mixtureobtained was stirred at room temperature for 2 days. A saturated aqueousNaHCO3 solution was then added and the mixture was extracted twice withdiethyl ether. The organic phase was washed with brine, dried oversodium sulphate and concentrated under vacuum to give 300 mg of crude.This was purified by flash chromatography (Biotage KP-Sil 25 g SNAPcolumn, eluant cyclohexane/ethyl acetate from 90/10 to 20/80 in 12CV) togive 106 mg of the title compound as a brown solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 8.13-8.18 (2H, m), 7.06 (1H, t), 6.85(1H, d), 6.66 (1H, d), 6.45 (1H, d), 4.85 (1H, br.s), 4.48 (2H, s),1.90-2.11 (2H, m), 1.53 (3H, s), 1.48 (9H, s), 1.42-1.47 (2H, m), 0.97(3H, t), 0.81-0.86 (2H, m); UPLC: 1.19 min, 454 [M+H]+.

Intermediate 93

N¹-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-D-isovalinamide

To a solution of1,1-dimethylethyl[(1R)-1-methyl-1-({[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate(Intermediate 92, 106 mg) in dry dichloromethane (2 ml) at 0° C. TFA(0.360 ml, 4.67 mmol) was added. The mixture was stirred at thistemperature for 5 minutes, then allowed to warm up at room temperature.After 2 hours UPLC/MS showed the absence of the starting material andthe presence of the desired compound: toluene (5 ml) was added and themixture was concentrated under vacuum. The residue was loaded on a SCXcartridge (1 g), which was eluted with methanol and a 1M NH₃ solution inmethanol. The basic eluate was concentrated under vacuum to afford thetitle compound as a brown oil (68 mg) which was used in the followingexperiment without further purification.

¹H NMR (400 MHz, CDCl₃): δ ppm 9.93 (1H, s), 8.21-8.31 (2H, m), 7.06(1H, t), 6.85 (1H, d), 6.65 (1H, d), 6.45 (1H, d), 4.48 (2H, s),1.94-2.04 (1H, m), 1.59-1.69 (1H, m), 1.44-1.48 (2H, m), 1.43 (3H, s),0.95 (3H, t), 0.81-0.85 (2H, m); UPLC: 0.71 min, 354 [M+H]+.

Intermediate 94 methyl 3-{[(methyloxy)methyl]oxy}benzoate

In a 500 ml round-bottomed flask, under argon flush, methyl3-hydroxybenzoate (5 g, 32.9 mmol) was dissolved in dichloromethane (100ml) to give a white suspension. The reaction mixture was cooled at 0° C.At that temperature chloro(methyloxy)methane (2.75 ml, 36.1 mmol) andDIPEA (6.89 ml, 39.4 mmol) were added. The reaction mixture was stirredovernight. During that time, the reaction temperature was allowed toreach room temperature. The reaction mixture was then evaporated undervacuum to afford the crude product as a yellow oil which was purified bysilica gel chromatography (Biotage SP1 system, 50 g SNAP column) withCyclohexane/EtOAc as eluents (from 10/0 to 3/1 in 10 CV; then 3/1 for 5CV). The collected fractions afforded the title compound (5.088 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.76-7.68 (2H, m), 7.41-7.34 (1H, m),7.27-7.22 (1H, m), 5.24 (2H, s), 3.93 (3H, s), 3.51 (3H, s); UPLC_ipqc:0.92 min, 197 [M+H]+

Intermediate 95 (3-{[(methyloxy)methyl]oxy}phenyl)methanol

In a 250 ml round-bottomed flask, under argon flush, methyl3-{[(methyloxy)methyl]oxy}benzoate (Intermediate 94, 5.0875 g) wasdissolved in tetrahydrofuran (20 ml) to give a colourless solution. Thereaction mixture was cooled at 0° C. In those conditions, a solution ofLiAlH₄ in (1M) (25.9 ml, 25.9 mmol) was added dropwise and the reactionmixture was stirred at 0° C. After 45 min, the reaction mixture wasquenched with a 2M hydrochloric acid solution until pH ˜2 and dilutedwith 100 ml of dichloromethane. Phases were separated through a phaseseparator cartridge. The organic phase was evaporated under vacuum toafford the title compound (4.348 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.30 (1H, t), 7.10-7.05 (1H, m),7.04-7.00 (1H, m), 7.00-6.95 (1H, m), 5.20 (2H, s), 4.69 (2H, d), 3.50(3H, s), 1.80-1.70 (1H, m).

UPLC_ipqc: 0.63 min, 151 [M−OH]⁺

Intermediate 961-(2-(hydroxymethyl)-6-{[(methyloxy)methyl]oxy}phenyl)ethanol

Under argon flush, in a 2-necked 100 ml round-bottomed flask equippedwith a reflux condenser (flammed for 5 min under vacuum and then 3cycles of Ar/vacuum) (3-{[(methyloxy)methyl]oxy}phenyl)methanol(Intermediate 95, 1 g) was dissolved in hexane (20 ml) to give acolourless solution. N,N,N′,N′-tetramethyl-1,2-ethanediamine (1.872 mL,12.49 mmol) was added. To the obtained reaction mixture a solution ofBuLi (1.6M/hexane) (7.80 ml, 12.49 mmol) was added dropwise. Thereaction mixture was then heated at 60° C. and stirred in thoseconditions. After 5 hours stirring in those conditions, acetaldehyde(1.090 ml, 19.30 mmol) in 6 ml of dry hexane was added dropwise at −78°C. The reaction mixture was stirred at that temperature for 1 hour andthen warmed to room temperature. After overnight stirring at thattemperature, the reaction was quenched with an aqueous solution of 2Mhydrochloric acid and diluted with 100 ml of dichloromethane. The phaseswere separated through a phase separator cartridge. The organic phasewas evaporated under vacuum to afford the crude product which waspurified by silica gel chromatography (Biotage SP1 system, 25 g-SNAPSilica column) with Cyclohexane/EtOAc as eluents (from 5/1 to 1/1 in 15CV; then 1/1 for 5 CV). The collected fractions afforded(3-{[(methyloxy)methyl]oxy}phenyl)methanol (532.4 mg, recovered

Intermediate 95) and the title compound (184.7 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.20 (1H, t), 7.10 (1H, dd), 7.00 (1H,dd), 5.37 (1H, q), 5.26 (1H, d), 5.25 (1H, d), 4.83 (1H, d), 4.63 (1H,d), 3.51 (3H, s), 3.55 (1H, br. s.), 2.66 (1H, br. s.), 1.60 (3H, d);UPLC_ipqc: 0.60 min, 195 [M−OH]+

Intermediate 97

1-methyl-7-{[(methyloxy)methyl]oxy}-1,3-dihydro-2-benzofuran

Under argon flush, in a 8 ml vial1-(2-(hydroxymethyl)-6-{[(methyloxy)methyl]oxy}phenyl)ethanol(Intermediate 96, 50.3 mg) was dissolved in tetrahydrofuran (1 ml) togive a pale yellow solution. The reaction mixture was cooled at 0° C. Inthat conditions, a solution of BuLi (1.6M/hexane) (0.148 ml, 0.237 mmol)was added. The reaction mixture was stirred at 0° C. After 30 minutes,4-methyl benzenesulfonyl chloride (45.2 mg, 0.237 mmol) was added at 0°C. The reaction was stirred at that temperature. After 1 h, anadditional quantity of the BuLi solution (1.6M/hexane) (0.148 ml, 0.237mmol) was added at 0° C. The reaction mixture was stirred at 0° C. for 1hour, then at room temperature for 30 minutes. The reaction mixture wasthen quenched with 2 ml of 2M hydrochloric acid and diluted with 5 ml ofdichloromethane. The phases were separated through a phase separatorcartridge. The organic phase was evaporated under vacuum to afford thecrude product which was purified by silica gel chromatography (BiotageSP1 system, 10 g—SNAP Silica column) with Cyclohexane/EtOAc as eluents(from 1/0 to 3/1 in 10 CV; then 3/1 for 5 CV; then from 3/1 to 1/1 in 5CV; then 1/1 for 5 CV). The collected fraction afforded the titlecompound (25.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.25 (1H, t), 6.99 (1H, d), 6.88 (1H, d),5.53-5.43 (1H, m), 5.26 (1H, d), 5.23 (1H, d), 5.20 (1H, dd), 5.10-5.02(1H, m), 3.51 (3H, s), 1.57 (3H, d); UPLC_ipqc: 0.92 min, 195 [M+H]+

And also the corresponding deprotected phenol (Intermediate 98, 11.1mg). ¹H NMR (400 MHz, CDCl₃): 6 ppm 7.17-7.11 (1H, m), 6.79 (1H, d),6.63 (1H, d), 5.55-5.40 (1H, m), 5.20-5.14 (2H, m), 5.04 (1H, d), 1.57(3H, d); UPLC_ipqc: 0.66 min, 149 [M−H]−

Intermediate 98 3-methyl-1,3-dihydro-2-benzofuran-4-ol

In a 8 ml vial1-methyl-7-{[(methyloxy)methyl]oxy}-1,3-dihydro-2-benzofuran(Intermediate 97, 25.2 mg) was dissolved in methanol (1 ml) to give acolourless solution. A solution of HCl (2M/H₂O) (0.259 ml, 0.519 mmol)was added. The reaction mixture was shaken at 80° C. After 30 minutes,the reaction mixture was diluted with 10 ml of dichloromethane. Thephases were separated through a phase separator cartridge. The organicphase was mixed with the fraction obtained before and evaporated undervacuum to afford a residue which was purified by silica gelchromatography (Biotage SP1 system, 10 g-SNAP Silica column) withCyclohexane/EtOAc as eluents (from 100/0 to 3/1 in 10 CV; then 3/1 for 5CV; then from 3/1 to 1:1 in 5 CV; then 1:1 for 5 CV). The collectedfractions afforded the title compound (24 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.14 (1H, t), 6.79 (1H, d), 6.64 (1H, d),5.55-5.42 (1H, m), 5.23 (1H, s), 5.17 (1H, dd), 5.04 (1H, d), 1.57 (3H,d); UPLC_ipqc: 0.66 min, 149 [M−H]−

Intermediate 992-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-5-nitropyridine

In a microwave vial 3-methyl-1,3-dihydro-2-benzofuran-4-ol (Intermediate98, 24 mg) was dissolved in N,N-dimethylformamide (1.5 ml) to give apale yellow solution. 2-chloro-5-nitropyridine (24.07 mg, 0.152 mmol)and potassium carbonate (62.9 mg, 0.455 mmol) were added. The reactionvessel was sealed and heated in a microwave Biotage Initiator at 110° C.for 1 hour. After cooling, the reaction mixture was quenched with 5 mlof water and diluted with 25 ml of Et₂O. The organic phase was washedwith water (3×10 mL) and the phases were separated. The organic phasewas passed through a phase separator cartridge and evaporated undervacuum to give the title compound (38.3 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, d), 8.53 (1H, dd), 7.39 (1H,t), 7.18 (1H, d), 7.10 (1H, d), 7.04 (1H, d), 5.37-5.26 (1H, m),5.26-5.18 (1H, m), 5.12 (1H, d), 1.41 (3H, d); UPLC_B: 0.84 min, 273.[M+H]+

Intermediate 1006-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinamine

In a 50 ml round-bottomed flask2-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-5-nitropyridine(Intermediate 99, 38.3 mg) was dissolved in ethanol (10 ml) to give apale yellow solution. Pd/C (14.22 mg, 0.013 mmol) and hydrazine hydrate(0.026 ml, 0.267 mmol) were added. The reaction mixture was stirred at90° C. After 45 minutes, the reaction mixture was filtered and theorganic phase was evaporated under vacuum to afford the title compound(32 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.70 (1H, d), 7.25 (1H, t), 7.09 (1H,dd), 6.99 (1H, d), 6.88 (1H, d), 6.76 (1H, d), 5.28-5.39 (1H, m), 5.18(1H, dd), 5.07 (1H, d), 3.53 (2H, br. s.), 1.47 (3H, d); UPLC_B: 0.64min, 243 [M+H]+

Intermediate 101 1,1-dimethylethyl{(1R)-1-[({6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

In a 8 ml vial (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoicacid (32.2 mg, 0.158 mmol) was dissolved in N,N-dimethylformamide (0.5ml) to give a colourless solution. DIPEA (0.035 ml, 0.198 mmol) and HATU(60.3 mg, 0.158 mmol) were added. The reaction mixture was stirred atroom temperature for 30 minutes.6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 100, 32 mg) was dissolved in 1.5 ml of DMF and theobtained solution was added to the reaction mixture. It was shaken at60° C. After 2 hours, no reaction occurred. Additional 0.5 ml of asolution [obtained dissolving in 0.5 ml of DMF(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (32.2 mg,0.158 mmol), DIPEA (0.035 mL, 0.198 mmol) and HATU (60.3 mg, 0.158mmol)] was added to the reaction mixture. It was shaken at 60° C. overweekend. After that time, only traces of the title compound along withthe Intermediate 100 was detected. The reaction mixture was evaporatedunder vacuum using the Vaportec V10 to give the crude product which waspurified by silica gel chromatography (Biotage SP1 system, 10 g-SNAPSilica column) with Cyclohexane/EtOAc as eluents (from 3/1 to 1/1 in 10CV; then 1/1 for 10 CV). The collected fractions afforded 4.9 mg of thetitle compound mixed with the amino acid and 61 mg of a mixture ofstarting material (Intermediate 100) and the amino acid.

This was dissolved in 10 ml of dichloromethane and washed with 10 ml ofsaturated aqueous solution of sodium bicarbonate. The phases wereseparated through a phase separator cartridge. The organic phases wereevaporated under vacuum to afford 64 mg of the same mixture. This latterwas dissolved in 1.0 ml of DMF and added to a stirring DMF solution (0.5ml) of (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid(61.4 mg, 0.302 mmol), DIPEA (0.066 mL, 0.378 mmol) and TBTU (97 mg,0.302 mmol). The obtained mixture was warmed at 60° C. and was shaken.After 1 h 30 min the reaction mixture was evaporated under vacuum usingthe Vaportec V10 to give the crude product as a yellow oil which wasdissolved in 10 ml of EtOAc and quenched with 10 ml of a saturatedaqueous solution of sodium bicarbonate. The aqueous phase was extractedwith EtOAc (3×10 ml). The collected organic phases were dried using ahydrophobic frit, mixed with the title compound obtained before andpurified by silica gel chromatography (Biotage SP1 system, 10 g-SNAPSilica column) with Cyclohexane/EtOAc as eluents (from 3/1 to 1/1 in 10CV; then 1:1 for 10 CV). The collected fractions afforded the titlecompound, as a 1:1 mixture of diastereoisomers (62.3 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.20-8.16 (1H, m), 8.15-8.08 (1H, m),7.31 (1H, t), 7.06 (1H, d), 6.95 (1H, d), 6.89 (1H, d), 5.37-5.25 (1H,m), 5.20 (1H, dd), 5.15-5.02 (2H, m), 4.23-4.08 (1H, m), 2.03-1.81 (2H,m), 1.47 (9H, s), 1.43 (3H, d), 1.08-0.99 (3H, m). UPLC_ipqc: 1.05 min,428 [M+H]+

Intermediate 102(2R)-2-amino-N-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide

In a 25 ml round-bottomed flask 1,1-dimethylethyl{(1R)-1-[({6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 101, 62.3 mg) was dissolved in dichloromethane (3 ml) togive a pale yellow solution. The reaction mixture was cooled at 0° C. Atthat temperature TFA (0.5 ml, 6.49 mmol) was added dropwise to thereaction mixture. The reaction mixture was stirred at 0° C. After 1hour, the reaction mixture was evaporated under vacuum to afford thecrude product which was charged on a 2 g SCX cartridge. It was thenflushed with 50 ml of MeOH followed by 25 ml of a 2M solution of ammoniain MeOH. The ammonia eluate was evaporated under vacuum to afford 34.4mg of a yellow oil which was a mixture of the title compound and thedeprotected amino acid. This mixture was dissolved in 20 ml of Et2O andwas washed with a saturated aqueous solution of NaHCO₃ (3×10 ml). Theorganic phase was dried using a phase separator cartridge and evaporatedunder vacuum to afford the title compound as a 1:1 mixture ofdiastereoisomers (24.4 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.58 (1H, br. s.), 8.32-8.25 (1H, m),8.23 (1H, t), 7.32 (1H, t), 7.08 (1H, d), 6.98 (1H, d), 6.94 (1H, d),5.40-5.29 (1H, m), 5.22 (1H, dd), 5.11 (1H, d), 3.49 (1H, dd), 2.11-1.94(1H, m), 1.85-1.60 (3H, m), 1.46 (3H, d), 1.12-1.01 (3H, m); UPLC_ipqc:0.64 min, 328 [M+H]+

Intermediate 103(1,1-dimethylethyl)(dimethyl){[(3-{[(methyloxy)methyl]oxy}phenyl)methyl]oxy}silane

In a 100 ml round-bottomed flask(3-{[(methyloxy)methyl]oxy}phenyl)methanol (Intermediate 95, 3.5 g) wasdissolved in dichloromethane (20 ml) to give a colourless solution.1H-imidazole (1.700 g, 24.97 mmol) andchloro(1,1-dimethylethyl)dimethylsilane (3.64 g, 24.14 mmol) were added.The reaction mixture immediately became a white suspension and wasstirred at room temperature. After overnight stirring the reaction wascompleted. The reaction mixture was then quenched with 10 ml of waterand diluted with 10 ml of dichloromethane. The phases were separatedthrough a separating funnel. The organic phase was dried using ahydrophobic frit and evaporated under vacuum to give 6.0082 g of thecrude product as a colourless oil which was purified via Biotage SP1(with cyclohexane/EtOAc as eluents from 1:0 to 5:1 in 10 CV; then 5:1for 5 CV; then from 5:1 to 1:1 in 5 CV; 100 g SNAP Silica column). Twofractions of the title compound were collected: 1.70 g of a colourlessoil (purity: 93%) and 3.70 g of a colourless oil (purity: 98%).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.26 (1H, t), 7.04-7.08 (1H, m), 6.96-7.01(1H, m), 6.91-6.96 (1H, m), 5.20 (2H, s), 4.75 (2H, s), 3.50 (3H, s),0.92-1.02 (9H, m), 0.12 (6H, s). UPLC-MS_ipqc: 1.49 min, 281 [M−H]−.

Intermediate 1041-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)-1-propanol

Under argon flush, in a 2-necked 100 ml round-bottomed flask equippedwith a reflux condenser (flammed for 5 minutes under vacuum and then 3cycles of Ar/vacuum)(1,1-dimethylethyl)(dimethyl){[(3-{[(methyloxy)methyl]oxy}phenyl)methyl]oxy}silane(Intermediate 103, 0.2 g) was dissolved in hexane (2 ml) to give acolourless solution. A 1.6M/Hexane solution of BuLi (0.487 ml, 0.779mmol) was added dropwise. The reaction mixture was stirred at roomtemperature. After 2 hours stirring in those conditions, to the paleyellow reaction mixture propanal (0.061 mL, 0.850 mmol) was added at 0°C. After 1 h 30 min, the reaction was quenched with 2M hydrochloric aciduntil pH ˜2 and diluted with 25 ml of dichloromethane. The phases wereseparated through a phase separator cartridge. The organic phase wasevaporated under vacuum affording the crude product as a green/grey oilthat was purified via Biotage SP1 (with Cyclohexane/EtOAc as eluentsfrom 1:0 to 5:1 in 5 CV; then 5:1 for 5 CV; then from 5:1 to 3:1 in 5CV; then 3:1 for 5 CV; 10 g SNAP Silica column.) This afforded the titlecompound as a pale yellow oil (132.1 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.18 (1H, t), 7.02-7.12 (2H, m), 5.26 (1H,d), 5.25 (1H, d), 4.78-4.89 (2H, m), 4.75 (1H, d), 3.63 (1H, d), 3.51(3H, s), 1.89-2.06 (1H, m), 1.74-1.88 (1H, m), 1.02 (3H, t), 0.94 (9H,s), 0.12 (3H, s), 0.10 (3H, s). UPLC-MS_ipqc: 1.45 min, 363 [M+Na]+.

Intermediate 1051-(2-(hydroxymethyl)-6-{[(methyloxy)methyl]oxy}phenyl)-1-propanol

In a 50 ml round-bottomed flask1-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)-1-propanol(Intermediate 104, 132.1 mg) was dissolved in Tetrahydrofuran (2 ml) togive a pale yellow solution. 1M/THF sol of TBAF (0.388 ml, 0.388 mmol)was added. The reaction mixture was stirred at room temperature. Afterover night stirring, the reaction was completed. The reaction mixturewas evaporated under vacuum to give the crude product as a pale yellowoil which was purified via Biotage SP1 (with Cyclohexane/EtOAc aseluents from 3:1 to 2:1 in 10 CV; then 2:1 for 5 CV; then from 2:1 to2:1 in 5 CV; then 1:1 for 5 CV; 25 g SNAP Silica column). The collectedfractions afforded a residue which was purified again in the sameconditions to afford the title compound as a pale yellow oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.16-7.24 (1H, m), 7.10 (1H, dd), 7.02(1H, dd), 5.23 (2H, dd), 5.08 (1H, dd), 4.81 (1H, d), 4.61 (1H, d), 3.50(3H, s), 2.41-3.05 (2H, m), 1.91-2.07 (1H, m), 1.82 (1H, s), 1.00 (3H,t). UPLC-MS_ipqc: 0.70 min, 249 [M+Na]+.

Intermediate 106 3-ethyl-1,3-dihydro-2-benzofuran-4-ol

Under argon flush, in a 8 ml vial1-(2-(hydroxymethyl)-6-{[(methyloxy)methyl]oxy}phenyl)-1-propanol(Intermediate 105, 99 mg) was dissolved in tetrahydrofuran (2 ml) togive a pale yellow solution. The reaction mixture was cooled at 0° C. Inthose conditions, a solution of BuLi in hexane (1.6M, 0.246 ml, 0.394mmol) was added. The reaction mixture was stirred at 0° C. After 30minutes, 4-methylbenzenesulfonyl chloride (75 mg, 0.394 mmol) was addedat 0° C. The reaction was stirred at that temperature. After 1 hour,additional BuLi in hexane (1.6M, 0.246 ml, 0.394 mmol) was added at 0°C. The reaction mixture was stirred at 0° C. for 1 h, then at roomtemperature for 30 minutes. After that time the reaction mixture wasquenched with 2 ml of 2M hydrochloric acid and diluted with 5 ml ofdichloromethane. The phases were separated through a phase separatorcartridge. The organic phase was evaporated under vacuum to afford thecrude product as a pale yellow oil which was dissolved in methanol (3.00ml). HCl (0.788 ml, 1.575 mmol) was added thereto. The obtained reactionmixture was warmed at 80° C. and shaken. After 30 minutes, the reactionwas completed. The reaction mixture was quenched with 5 ml of water anddiluted with 25 ml of dichloromethane. The phases were separated througha phase separator cartridge. The organic phase was evaporated undervacuum to afford the crude product as a pale yellow oil which waspurified via Biotage SP1 (with Cyclohexane/EtOAc as eluents from 1:0 to3:1 in 10 CV; then 3:1 for 5 CV; then from 3:1 to 1:1 in 5 CV; then 1:1for 5 CV; 10 g SNAP Silica column). The collected fractions afforded thetitle compound as a colourless oil (49.7 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.14 (1H, t), 6.79 (1H, d), 6.64 (1H, d),5.51 (1H, br. s.), 5.37-5.45 (1H, m), 5.16 (1H, dd), 5.07 (1H, d),1.99-2.15 (1H, m), 1.78-1.92 (1H, m), 0.91-1.00 (3H, m). UPLC-MS_ipqc:0.77 min, 163 [M−H]−.

Intermediate 1072-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-5-nitropyridine

In a 0.5-2.0 ml micro-wave vial 3-ethyl-1,3-dihydro-2-benzofuran-4-ol(Intermediate 106, 49.7 mg) was dissolved in N,N-dimethylformamide (1.5ml) to give a pale yellow solution. 2-chloro-5-nitropyridine (48.0 mg,0.303 mmol) and potassium carbonate (125 mg, 0.908 mmol) were added. Thereaction vessel was sealed and heated in micro-wave Biotage Initiator at110° C. for 1 hour. After cooling, the reaction completed. The reactionmixture was then quenched with 5 ml of water and diluted with 25 ml ofEt₂O. The aqueous phase was extracted with 3×10 ml of Et₂O. Thecollected organic phases were passed through a phase separator cartridgeand evaporated under vacuum to give the title compound as a pale yellowoil (97.6 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.05 (1H, d), 8.51 (1H, dd), 7.37 (1H, t),7.16 (1H, d), 7.06 (1H, d), 7.02 (1H, d), 5.16-5.25 (2H, m), 5.13 (1H,d), 1.77-1.90 (1H, m), 1.60-1.72 (1H, m), 0.82-0.91 (3H, m). UPLC_B:0.90 min, 287 [M+H]+.

Intermediate 1086-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinamine

In a 50 ml round-bottomed flask2-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-5-nitropyridine(Intermediate 107, 97.6 mg) was dissolved in ethanol (10 ml) to give apale yellow solution. Pd/C (26.1 mg, 0.245 mmol) and hydrazine hydrate(12.29 mg, 0.245 mmol) were added. The reaction mixture was stirred at90° C. After 45 minutes, the reaction was completed. The reactionmixture was filtered and the organic phase was evaporated under vacuumaffording the crude product which was charged on a 2 g SCX cartridge. Itwas then flushed with 15 ml of methanol followed by 15 ml of a 2Msolution of ammonia in methanol. The ammonia eluate was evaporated undervacuum affording nothing. The methanol eluate was then evaporated undervacuum to afford the title compound as a colourless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.70 (1H, d), 7.24 (1H, t), 7.09 (1H, dd),6.99 (1H, d), 6.87 (1H, d), 6.75 (1H, d), 5.21-5.29 (1H, m), 5.17 (1H,dd), 5.09 (1H, d), 3.57 (2H, br. s.), 1.88-2.04 (1H, m), 1.68-1.82 (1H,m), 0.89 (3H, t). UPLC_B: 0.71 min, 257 [M+H]+.

Intermediate 109 1,1-dimethylethyl{(1R)-1-[({6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

In a 8 ml vial6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 108, 10 mg) was dissolved in N,N-dimethylformamide (1 ml)to give a colourless solution. DIPEA (10.22 μl, 0.059 mmol),(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (9.52 mg,0.047 mmol) and, finally, TBTU (15.03 mg, 0.047 mmol) were added. Thereaction mixture was shaken at 60° C. After overnight shaking, thereaction was completed. The reaction mixture was evaporated under vacuumto give the crude product as a yellow oil which was purified via BiotageSP1 (with Cyclohexane/EtOAc as eluents from 2:1 to 1:1 in 10 CV; then2:1 for 5 CV; 10 g SNAP Silica column). The collected fractions affordedthe title compound as a colourless oil (12.7 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.40 (1H, br. s.), 8.15-8.18 (1H, m),8.08-8.14 (1H, m), 7.30 (1H, t), 7.05 (1H, d), 6.95 (1H, d), 6.88 (1H,d), 5.20-5.26 (1H, m), 5.17 (1H, dd), 5.10 (1H, d), 4.99 (1H, br. s.),4.05-4.19 (1H, m), 1.82-2.08 (2H, m), 1.64-1.79 (2H, m), 1.47 (9H, s),1.03 (3H, t), 0.88 (3H, t). UPLC_B: 0.91 min, 442 [M+H]+.

Intermediate 110(2R)-2-amino-N-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide

In a 50 ml round-bottomed flask 1,1-dimethylethyl{(1R)-1-[({6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 109, 12.7 mg) was dissolved in dichloromethane (3 ml) togive a colourless solution. The reaction mixture was cooled at 0° C. TFA(1.5 ml, 19.47 mmol) was added at that temperature. The reaction mixturewas stirred at 0° C. After 1 hour, the reaction was completed. Thereaction mixture was evaporated under vacuum affording the crude productwhich was charged on a 2 g SCX cartridge. It was then flushed with 15 mlof MeOH followed by 15 ml of a solution of ammonia in MeOH (2M). Theammonia eluate was evaporated under vacuum to afford the title compoundas a colorless oil (9.0 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.59 (1H, br. s.), 8.18-8.28 (2H, m), 7.30(1H, t), 7.05 (1H, d), 6.95 (1

H, d), 6.90 (1H, d), 5.21-5.26 (1H, m), 5.18 (1H, dd), 5.10 (1H, d),3.50 (1H, dd), 1.79-2.07 (4H, m), 1.62-1.78 (2H, m), 1.04 (3H, t), 0.88(3H, t); UPLC_B: 0.72 min, 342 [M+H]+.

Intermediate 111 3-methyl-5-(methyloxy)-2H-chromen-2-one

To a solution of 2-hydroxy-6-(methyloxy)benzaldehyde (3 g, 19.72 mmol)in dry N,N-dimethylformamide (30 ml), propanoic anhydride (12.98 ml, 101mmol) and K₂CO₃ (3.00 g, 21.69 mmol) were added and the reaction mixturewas warmed to 70° C. At this temperature water (0.036 ml, 1.972 mmol)was added and the reaction mixture was warmed to 120° C. and stirredovernight under nitrogen. The reaction was then quenched with 60 ml ofwater. A precipitate was formed and the crude material was filtered, thesolid dissolved in DCM/water and the two phases were separated through aphase separator cartridge. The organic phase was evaporated under vacuumaffording the title compound as a white solid (3.25 g, 85% yield).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.91 (1H, s), 7.37 (1H, t), 6.92 (1H, d),6.71 (1H, d), 3.94 (3H, s), 2.22 (3H, s); UPLC_ipqc: 0.95 min, 191[M+H]+.

Intermediate 112 5-hydroxy-3-methyl-2H-chromen-2-one

To a solution of 3-methyl-5-(methyloxy)-2H-chromen-2-one (Intermediate111, 2.5 g) in dry dichloromethane (45 ml) cooled to 0° C. was addedBBr3 (39.4 ml, 39.4 mmol). The reaction mixture was warmed to roomtemperature and stirred at room temperature overnight under nitrogen.The reaction was then cooled to 0° C. and quenched with ice. Theobtained mixture was diluted with diethyl ether and the two phasesobtained were separated through a separating funnel. The aqueous phasewas back-extracted with diethyl ether. The collected organic phases weredried over sodium sulphate, filtered and evaporated under vacuum to givethe title compound as a light brown solid (2.225 g).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.92 (1H, s), 7.30 (1H, t), 6.93 (1H, d),6.67 (1H, d), 5.54 (1H, s), 2.26 (3H, s); UPLC_ipqc: 0.70 min, 177[M+H]+.

Intermediate 113 3-methyl-5-{[(methyloxy)methyl]oxy}-2H-chromen-2-one

To a solution of 5-hydroxy-3-methyl-2H-chromen-2-one (Intermediate 112,2.225 g) in dry N,N-Dimethylformamide (60 ml) cooled to 0° C. was addedsodium hydride (60%, 0.532 g, 13.89 mmol) followed by the addition ofchloro(methyloxy)methane (1.919 ml, 25.3 mmol). The reaction mixture waswarmed to room temperature and stirred for 30 minutes under nitrogen. Itwas then quenched by the addition of a saturated solution of NH₄Cl, theproduct was extracted with diethyl ether, washed with brine, dried oversodium sulphate, filtered and concentrated under vacuum. The residueobtained was purified by silica gel chromatography (Biotage system, witha gradient from pure cyclohexane to cyclohexane/ethyl acetate 5/1) togive the title compound as a white solid (2.15 g). ¹H NMR (400 MHz,CDCl₃): δ ppm 7.94 (1H, s), 7.38 (1H, t), 6.93-7.05 (2H, m), 5.33 (2H,s), 3.55 (3H, s), 2.26 (3H, s); UPLC_ipqc: 0.94 min, 221 [M+H]+.

Intermediate 114 3:1 mixture of2-[3-hydroxy-2-methyl-1-propen-1-yl]-3-{[(methyloxy)methyl]oxy}phenoland 2-(3-hydroxy-2-methylpropyl)-3-{[(methyloxy)methyl]oxy}phenol

In a 50 ml round-bottomed flask3-methyl-5-{[(methyloxy)methyl]oxy}-2H-chromen-2-one (Intermediate 113,461.1 mg) was dissolved in tetrahydrofuran (3 ml) to give a colourlesssolution. The reaction mixture was cooled at 0° C. A solution of LiAlH₄(1M/THF, 4.19 ml, 4.19 mmol) was then added dropwise. The reactionmixture was stirred at 0° C. After 30 minutes, the reaction mixture wasquenched with 5 ml of hydrochloric acid (2M) and diluted with 10 ml ofdichloromethane. Phases were separated through a phase separatorcartridge. The organic phase was evaporated under vacuum to afford a 3:1mixture of2-[3-hydroxy-2-methyl-1-propen-1-yl]-3-{[(methyloxy)methyl]oxy}phenoland 2-(3-hydroxy-2-methylpropyl)-3-{[(methyloxy)methyl]oxy}phenol (548mg) as a pale yellow oil.

¹H NMR (400 MHz, CDCl₃) major component of the mixture δ ppm 7.11 (1H,t), 6.67 (1H, d), 6.63 (1H, d), 6.08-6.16 (1H, m), 5.81 (1H, br. s.),5.15 (2H, s), 4.74 (1H, br. s.), 4.00 (2H, s), 3.47 (3H, s), 2.08 (3H,d); ¹H NMR (400 MHz, CDCl₃) minor component of the mixture δ ppm 7.05(1H, t), 6.97 (1H, br. s.), 6.66 (1H, d), 6.58 (1H, d), 5.19 (2H, s),3.51-3.59 (1H, m), 3.49 (3H, s), 3.41-3.49 (1H, m), 2.78 (1H, dd), 2.66(1H, dd), 2.13 (1H, br. s.), 1.94-2.07 (1H, m), 1.09 (3H, d);UPLC-MS_ipqc: major component of the mixture: 0.70 min, 223 [M−H]−;minor component of the mixture: 0.78 min, 225 [M−H]−.

Intermediate 1152-(3-hydroxy-2-methylpropyl)-3-{[(methyloxy)methyl]oxy}phenol

In a 100 ml round-bottomed flask a 3:1 mixture of2-[3-hydroxy-2-methyl-1-propen-1-yl]-3-{[(methyloxy)methyl]oxy}phenoland 2-(3-hydroxy-2-methylpropyl)-3-{[(methyloxy)methyl]oxy}phenol

(Intermediate 114, 548 mg) was dissolved in methanol (10 ml) to give acolourless solution. Three cycles of vacuum/N₂ were performed then Pd/C(129 mg, 0.122 mmol) was added to the reaction mixture. Again threecycles of vacuum/N₂ were performed before three cycles of vacuum/H₂.Finally, the reaction mixture was allowed to stir at room temperature inH₂ atmosphere (no pressure). The reaction mixture was stirred at roomtemperature. After 1 h 30 min the reaction was completed. The reactionmixture was filtered over a celite pad. The filtrate was evaporatedunder vacuum affording the crude product which was purified on theBiotage SP1 system with Cyclohexane/EtOAc as eluents from 1:0 to 5:1 in10 CV; then 5:1 for 5 CV; then from 5:1 to 3:1 in 5 CV; then 3:1 for 5CV; then from 3:1 to 1:1 in 5 CV; then 1:1 for 5 CV (25 g SNAP Silicacolumn). The collected fractions afforded the title compound as acolourless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.05 (1H, t), 6.97 (1H, br. s.), 6.66 (1H,d), 6.58 (1H, d), 5.19 (2H, s), 3.51-3.59 (1H, m), 3.49 (3H, s),3.41-3.49 (1H, m), 2.78 (1H, dd), 2.66 (1H, dd), 2.13 (1H, br. s.),1.94-2.07 (1H, m), 1.09 (3H, d). UPLC-MS_ipqc: 0.79 min, 225 [M−H]−.

Intermediate 1163-methyl-5-{[(methyloxy)methyl]oxy}-3,4-dihydro-2H-chromene

In a 50 ml round-bottomed flask under argon,2-(3-hydroxy-2-methylpropyl)-3-{[(methyloxy)methyl]oxy}phenol(Intermediate 115, 377.9 mg) was dissolved in tetrahydrofuran (5 ml) togive a colourless solution. TEA (0.409 ml, 2.93 mmol) was added and thereaction mixture was cooled at 0° C. At that temperature methanesulfonylchloride (0.124 ml, 1.591 mmol) was added. The reaction mixture wasstirred at 0° C. After 45 minutes, additional methanesulfonyl chloride(0.124 ml, 1.591 mmol) was added. After additional 45 minutes, potassium2-methyl-2-propanolate (422 mg, 3.76 mmol) was added. After 15 minutesfrom this latter addition, additional potassium 2-methyl-2-propanolate(422 mg, 3.76 mmol) was added. After 15 minutes the reaction wascompleted. The reaction was then quenched with 10 ml of a saturatedaqueous solution of NH₄Cl, acidified until pH˜2 with 2M hydrochloricacid and diluted with 25 ml of dichloromethane. Phases were separatedthrough a phase separator cartridge. The organic phase was evaporatedunder vacuum to afford the title compound.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.04 (1H, t), 6.63 (1H, d), 6.53 (1H, d),5.21 (2H, s), 4.13-4.19 (1H, m), 3.60-3.71 (1H, m), 3.50 (3H, s),2.84-2.95 (1H, m), 2.19-2.30 (1H, m), 2.04-2.18 (1H, m), 1.08 (3H, d);UPLC-MS_ipqc: 1.16 min, 209 [M+H]+.

Intermediate 117 3-methyl-3,4-dihydro-2H-chromen-5-ol

In a 8 ml vial3-methyl-5-{[(methyloxy)methyl]oxy}-3,4-dihydro-2H-chromene(Intermediate 116, 103.6 mg) was dissolved in methanol (3 ml) to give acolourless solution. A 2M/H₂O solution of HCl (0.224 ml, 0.448 mmol) wasadded. The reaction mixture was shaken at 60° C. After 2 h 30 min, thereaction mixture was diluted with 10 ml of dichloromethane. The phaseswere separated through a phase separator cartridge. The organic phasewas evaporated under vacuum to afford the crude product which waspurified via Biotage SP1 (with Cyclohexane/EtOAc as eluents from 1:0 to5:1 in 10 CV; then 5:1 for 5 CV; then from 5:1 to 3:1 in 5 CV; (10 gSNAP Silica column). The collected fractions afforded the title compoundas a white solid (53.0 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 6.96 (1H, t), 6.46 (1H, d), 6.35 (1H, dd),4.79 (1H, s), 4.10-4.22 (1H, m), 3.61-3.73 (1H, m), 2.78-2.90 (1H, m),2.19-2.30 (1H, m), 2.16 (1H, d), 1.09 (3H, d). UPLC-MS_ipqc: 0.90 min,165 [M+H]+.

Intermediate 1182-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-nitropyridine

In a microwave vial, 3-methyl-3,4-dihydro-2H-chromen-5-ol (Intermediate117, 53 mg), potassium carbonate (134 mg, 0.968 mmol) and2-chloro-5-nitropyridine (51.2 mg, 0.323 mmol) were dissolved inN,N-dimethylformamide (2 ml) to give a light brown solution. Thereaction vessel was sealed and heated in Biotage Initiator at 110° C.for 1 hour. After cooling, the reaction mixture was quenched with 5 mlof water and diluted with 10 ml of Et₂O. Phases were separated by aseparating funnel. The aqueous phase was extracted with 3×10 ml of Et₂O.The collected organic phase were dried using a hydrophobic frit andevaporated under vacuum to give the title compound as a brown oil (181mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.06 (1H, d), 8.48 (1H, dd), 7.17 (1H, t),7.02 (1H, d), 6.80 (1H, d), 6.66 (1H, dd), 4.16-4.24 (1H, m), 3.63-3.75(1H, m), 2.62-2.74 (1H, m), 2.04-2.18 (2H, m), 1.00 (3H, d). UPLC_B:0.95 min, 287 [M+H]+.

Intermediate 1196-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinamine

In a 50 ml round-bottomed flask2-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-nitropyridine(Intermediate 118, 181 mg) was dissolved in ethanol (10 ml) to give apale yellow solution. Pd/C (33.0 mg, 0.031 mmol) and hydrazine hydrate(0.030 ml, 0.310 mmol) were added. The reaction mixture was stirred at90° C. After 1 hour, the reaction mixture was filtered and evaporatedunder vacuum to give the title compound (131.4 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.72 (1H, d), 7.02-7.13 (2H, m), 6.72 (1H,d), 6.65 (1H, d), 6.51 (1H, dd), 4.11-4.20 (1H, m), 3.67 (1H, s), 3.52(2H, br. s.), 2.81-2.89 (1H, m), 2.16-2.25 (1H, m), 2.05-2.16 (1H, m),1.02 (3H, d). UPLC_B: 0.75 min, 257 [M+H]+.

Intermediate 1201,1-dimethylethyl[1,1-dimethyl-2-({6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}amino)-2-oxoethyl]carbamate

In a 8 ml vial6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinamine(Intermediate 119, 131.4 mg) was dissolved in N,N-dimethylformamide (2ml) to give a pale yellow solution. DIPEA (0.215 ml, 1.230 mmol),N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (188 mg, 0.923mmol) and HATU (351 mg, 0.923 mmol) were added. The reaction mixture wasshaken at 60° C. After 3 hours, the reaction mixture was evaporatedunder vacuum to give the crude product which was purified via BiotageSP1 (with Cyclohexane/EtOAc as eluents from 3:1 to 1:2 in 15 CV; then1:2 for 5 CV; 25 g SNAP Silica column.) The collected fractions affordedthe title as a white solid (104.0 mg).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.58 (1H, br. s.), 8.29 (1H, br. s.),7.97-8.13 (1H, m), 7.08 (1H, t), 7.00 (1H, br. s.), 6.93 (1H, d), 6.63(1H, dd), 6.53 (1H, dd), 4.08-4.17 (1H, m), 3.62 (1H, dd), 2.58-2.71(1H, m), 1.92-2.13 (2H, m), 1.36 (15H, br. s.), 0.93 (3H, d). UPLC_B:0.91 min, 442 [M+H]+.

Intermediate 1212-methyl-N1-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}alaninamide

In a 50 ml round-bottomed flask1,1-dimethylethyl[1,1-dimethyl-2-({6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}amino)-2-oxoethyl]carbamate(Intermediate 120, 104.0 mg) was dissolved in dichloromethane (6 ml) togive a pale yellow solution. The reaction mixture was cooled at 0° C.and TFA (2 ml, 26.0 mmol) was added. The reaction mixture was stirred at0° C. After 2 h 30 min, the mixture was evaporated under vacuum to givethe crude product which was charged on a 5 g SCX cartridge. It was thenflushed with 40 ml of methanol followed by 40 ml of a 2M solution ofammonia in methanol. The ammonia eluate was evaporated under vacuum togive the title compound as a colourless oil (72.6 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.90 (1H, br. s.), 8.17-8.26 (2H, m), 7.10(1H, t), 6.84 (1H, d), 6.70 (1H, d), 6.56-6.62 (1H, m), 4.12-4.22 (1H,m), 3.67 (1H, t), 2.70-2.85 (1H, m), 2.13 (2H, s), 1.47 (6H, s), 1.00(3H, d), NH2 Missed. UPLC_B: 0.78 min, 342 [M+H]+.

Intermediate 1221a-methyl-7-{[(methyloxy)methyl]oxy}-1a,7b-dihydrocyclopropa[c]chromen-2(1H)-one

To a solution of trimethylsulfoxonium iodide (4.52 g, 20.55 mmol) in drydimethyl sulfoxide (50 ml) stirred under nitrogen at room temperaturewas added neat sodium hydride (60%, 0.822 g, 20.55 mmol). The reactionmixture was stirred at room temperature for 1 hour before adding3-methyl-5-{[(methyloxy)methyl]oxy}-2H-chromen-2-one (Intermediate 113,1.81 g) dissolved into 15 ml of DMSO. The reaction mixture turned yellowand was heated and stirred at 100° C. for 4 h. The reaction was thenworked up by the addition of a saturated solution of NH₄Cl and extractedwith diethyl ether, washed with brine, dried over sodium sulphate,filtered and concentrated under vacuum. The residue obtained waspurified by silica gel chromatography (Biotage system, with a gradientfrom pure cyclohexane to cyclohexane/ethyl acetate 5/1) to give thetitle compound as a white solid (360 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.10 (1H, t), 6.88 (1H, d), 6.67 (1H, d),5.26 (2H, s), 3.53 (3H, s), 2.71 (1H, dd), 1.58 (1H, dd), 1.44 (3H, s),1.08 (1H, t); UPLC_ipqc: 0.96 min, 235 [M+H]+.

Intermediate 1232-[2-(hydroxymethyl)-2-methylcyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol

To a solution of1a-methyl-7-{[(methyloxy)methyl]oxy}-1a,7b-dihydrocyclopropa[c]chromen-2(1H)-one(Intermediate 122, 360 mg) in dry tetrahydrofuran (15 ml) stirred undernitrogen at 0° C. was added a solution of lithium aluminium hydride(1.0M in THF, 1.537 ml, 1.537 mmol) and the reaction mixture was stirredat that temperature for 20 minutes. The reaction was then diluted withTHF (20 ml) and quenched with the addition of Na₂SO₄.10H₂O (10 eq)leaving the mixture under stirring for 30 minutes. The reaction wasdiluted with ethyl acetate, dried over Na₂SO₄, filtered and concentratedunder vacuum. The residue obtained was purified by silica gelchromatography (Companion system, with a gradient from cyclohexane/ethylacetate 5/1 to cyclohexane/ethyl acetate 1/1) to give the title compoundas a white solid (307 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.40 (1H, br. s.), 7.06 (1H, t), 6.67(1H, d), 6.59 (1H, d), 5.12-5.27 (2H, m), 3.57-3.68 (1H, m), 3.53 (3H,s), 3.42-3.51 (1H, m), 2.98 (1H, d), 1.54 (1H, dd), 1.41 (3H, s), 1.05(1H, dd), 0.86 (1H, dd); UPLC_ipqc: 0.73 min, 237 [M−H]+.

Intermediate 1241a-methyl-7-{[(methyloxy)methyl]oxy}-1,1a,2,7b-tetrahydrocyclopropa[c]chromene

To a solution of2-[2-(hydroxymethyl)-2-methylcyclopropyl]-3-{[(methyloxy)methyl]oxy}phenol(Intermediate 123, 307 mg) in dry tetrahydrofuran (10 ml) were addedtriphenylphosphine (338 mg, 1.288 mmol) andbis(1-methylethyl)(E)-1,2-diazenedicarboxylate (261 mg, 1.288 mmol). Thereaction turned yellow and was stirred for 20 minutes at roomtemperature under nitrogen. The solvent was then evaporated under vacuumto give the crude product as pale yellow oil. The residue obtained waspurified by silica gel chromatography (Companion system, with a gradientfrom pure Cyclohexane to cyclohexane/ethyl acetate 10/1) to give thetitle compound as a white solid (280 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 6.97 (1H, t), 6.73 (1H, d), 6.53 (1H, d),5.15-5.30 (2H, m), 4.17 (1H, d), 3.67 (1H, d), 3.54 (3H, s), 2.15 (1H,dd), 1.27 (3H, s), 1.20 (1H, t), 0.94 (1H, dd); UPLC_ipqc: 1.15 min, 221[M+H]+.

Intermediate 125 1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-ol

To a solution of1a-methyl-7-{[(methyloxy)methyl]oxy}-1,1a,2,7b-tetrahydrocyclopropa[c]chromene(Intermediate 124, 280 mg) in methanol (16 ml) a 2.0 M aqueous solutionof HCl (1.271 ml, 2.54 mmol) was added. The reaction mixture was stirredat 50° C. for 2 hours and then at room temperature overnight at whichtime the reaction still showed some unreacted starting material,therefore more 2.0 M HCl was added (2 eq.) and stirring was continued at50° C. for 2 hours. The reaction mixture was then quenched with waterand diluted and extracted with dichloromethane, washed with brine, driedover sodium sulphate, filtered and concentrated under vacuum. Theresidue obtained was purified by silica gel chromatography (Companionsystem, with a gradient from pure cyclohexane to cyclohexane/ethylacetate 5/1) to give the title compound as a white solid (192 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 6.91 (1H, t), 6.40-6.53 (2H, m), 5.00(1H, s), 4.18 (1H, d), 3.72 (1H, d), 2.00 (1H, dd), 1.29 (3H, s), 1.20(1H, t), 0.98 (1H, dd); UPLC_ipqc: 0.91 min, 177 [M+H]+.

Intermediate 1262-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-5-nitropyridine

To a solution of 1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-ol(Intermediate 125, 190 mg) in dry N,N-dimethylformamide (10 ml) wereadded K₂CO₃ (447 mg, 3.23 mmol) and 2-chloro-5-nitropyridine (171 mg,1.078 mmol) to give a light brown solution. The reaction was heated at110° C. for 1 hour and then quenched with water, extracted with ethylacetate, washed with brine, dried over sodium sulphate, filtered andconcentrated under vacuum. The residue obtained was purified by silicagel chromatography (Companion system, with a gradient from pureCyclohexane to cyclohexane/ethyl acetate 5/1) to give the title compoundas a light brown solid (250 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.12 (1H, d), 8.51 (1H, dd), 7.12 (1H,t), 7.04 (1H, d), 6.81 (1H, d), 6.75 (1H, dd), 4.22 (1H, d), 3.74 (1H,d), 1.73 (1H, dd), 1.23 (1H, t), 1.21 (3H, s), 0.85 (1H, dd); UPLC_ipqc:1.21 min, 299 [M+H]+.

Intermediate 1276-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinamine

To a solution of2-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-5-nitropyridine(Intermediate 126, 250 mg) in tetrahydrofuran (6 ml)/water (3 ml) wereadded iron (234 mg, 4.19 mmol) and ammonium chloride (224 mg, 4.19mmol). The reaction mixture was stirred at room temperature overnight atwhich time UPLC showed partial formation of the target compound withsome hydroxyl-amine intermediate, therefore additional 3 equivalents ofammonium chloride and iron were added and the reaction was left stirringfor an other 5 hours. The iron was then filtered off over a pad ofcelite and the solution was diluted with an aqueous saturated solutionof NaHCO₃ (100 ml) and ethyl acetate (200 ml). Two phases were separatedand the aqueous layer was extracted with ethyl acetate. Organic layerswere combined, dried over sodium sulphate, filtered and evaporated. Theresidue was purified by silica gel chromatography (Companion system,with a gradient from cyclohexane/ethyl acetate 8:2 to cyclohexane/ethylacetate 1:1) to give the title compound as a light brown solid (150 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.75 (1H, d), 7.08 (1H, dd), 6.99 (1H,t), 6.73 (1H, d), 6.64 (1H, d), 6.59 (1H, d), 4.17 (1H, d), 3.70 (1H,d), 3.50 (2H, br. s.), 2.00 (1H, dd), 1.17-1.23 (4H, m), 0.85 (1H, dd);UPLC_ipqc: 0.88 min, 269 [M+H]+.

Intermediate 1281,1-dimethylethyl[1,1-dimethyl-2-({6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}amino)-2-oxoethyl]carbamate

To a solution of6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinamine(Intermediate 127, 150 mg) in dry N,N-dimethylformamide (10 ml) DIPEA(0.352 ml, 2.013 mmol),N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (307 mg, 1.509mmol) and HATU (574 mg, 1.509 mmol) were added. The reaction mixture washeated at 60° C. for 2.5 hours. The organic phase was washed withsaturated brine, extracted with diethyl ether, dried over sodiumsulphate, filtered and concentrated under vacuum. The residue waspurified by silica gel chromatography (Companion system, with a gradientfrom cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 1:1) togive the title compound as a white solid (135 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.17 (1H, d), 8.13 (1H, dd), 7.03 (1H,t), 6.86 (1H, d), 6.69 (2H, t), 5.86 (1H, br. s.), 4.89 (1H, br. s.),4.18 (1H, d), 3.71 (1H, d), 1.90 (1H, dd), 1.60 (6H, s), 1.45 (9H, s),1.15-1.23 (4H, m), 0.84 (1H, dd); UPLC_ipqc: 1.18 min, 454 [M+H]+.

Intermediate 1292-methyl-N¹-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}alaninamide

To a solution of1,1-dimethylethyl[1,1-dimethyl-2-({6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}amino)-2-oxoethyl]carbamate(Intermediate 128, 135 mg, 0.298 mmol) in dry dichloromethane (10 ml) at0° C. TFA (5 ml, 64.9 mmol) was slowly added and the reaction mixturewas stirred for 2 hours at the same temperature. The solvent and theexcess of TFA were evaporated and the residue was dissolved indichloromethane and a saturated solution of NaHCO₃ was slowly addedwhile the pH was allowed to reach 8. Two phases were separated and theorganic layer was dried over sodium sulphate, filtered and evaporatedaffording the title compound as a colourless oil used as it was withoutany further purification (105 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 9.94 (1H, br. s.), 8.19-8.30 (2H, m),7.03 (1H, t), 6.85 (1H, d), 6.70 (1H, d), 6.67 (1H, d), 4.18 (1H, d),3.71 (1H, d), 1.91 (1H, dd), 1.72 (2H, br. s.), 1.48 (6H, s), 1.15-1.23(4H, m), 0.84 (1H, dd); UPLC_ipqc: 0.76 min, 354 [M+1]+.

Intermediate 130 ethyl(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate

To a solution of(1,1-dimethylethyl)(dimethyl){[(3-{[(methyloxy)methyl]oxy}phenyl)methyl]oxy}silane(Intermediate 103, 3 g) in dry n-hexane (30 mL) a solution of BuLi inhexane (1.6M, 7.63 mL, 12.21 mmol) was added and the reaction mixturewas stirred for 2 hours at room temperature The reaction mixture wascooled to −78° C. and it was added (via cannulation) to a solution ofethyl chloro(oxo)acetate (1.780 mL, 15.93 mmol) in dry tetrahydrofuran(20 mL) at −78° C. The reaction was quenched with water (20 ml) andextracted with ethyl acetate (2×30 ml). The organic layer was dried oversodium sulphate, filtered and evaporated and the residue was purified byflash chromatography (Biotage system) on silica gel using a 100 g SNAPcolumn and cyclohexane to cyclohexane/ethyl acetate 8:2 as eluents. Thisafforded the title compound as a yellow pale oil (2.67 g).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.53 (1H, t), 7.20 (1H, d), 7.15 (1H,d), 5.21 (2H, s), 4.76 (2H, s), 4.27 (2H, q), 3.34 (3H, s), 1.27 (3H,t), 0.86 (9H, s), 0.05 (6H, s).

Intermediate 131 Dimethyltitanocene

To a suspension of dichlorotitanocene (8.3 g, 33.3 mmol) in dry toluene(100 mL) at −10° C. a solution of methyllithium in Et₂O (1.6M, 47.3 mL,76 mmol) was slowly added (20 minutes) and the reaction mixture wasstirred for 45 minutes at the same temperature. The reaction mixture wasadded (via cannulation) to a solution of ammonium chloride (1.2 g) inwater (24 ml) cooled to −10° C. Two phases were separated and theorganic layer was washed with cool water (3×20 ml) and brine (1×20 ml),dried over sodium sulphate filtered and concentrated under reducedpressure to 60 ml containing 9% w/w of dimethyltitanocene (5.04 g, 24mmol) in toluene.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 6.10 (10H, s), −0.11 (6H, s).

Intermediate 132Ethyl2-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)-2-propenoate

To a solution of ethyl(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)(oxo)acetate(Intermediate 130, 1.4 g) in dry toluene (8 ml) dimethyl titanocene 9%w/w in toluene (Intermediate 131, 30 ml) was added and the reactionmixture was stirred for 1.5 hours at 90° C. After cooling the reactionwas diluted with water (20 ml) and ethyl acetate (30 ml). Two phaseswere separated and the organic layer was dried over sodium sulphate,filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 100 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 9:1 as eluents. Thisafforded the title compound as a yellow pale oil (865 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.30 (1H, t), 7.12 (1H, d), 7.02 (1H,d), 6.46 (1H, d), 5.74 (1H, d), 5.12 (2H, s), 4.54 (2H, s), 4.12 (2H,q), 3.32 (3H, s), 1.16 (3H, t), 0.88 (9H, s), 0.04 (6H, s); UPLC_IPQC:1.54 min, 381 [M+H]+.

Intermediate 133 ethyl1-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate

To a solution of Trimethylsulfoxonium Iodide (816 mg, 3.71 mmol) in dryDimethyl Sulfoxide (10 mL)

NaH 60% disp in mineral oil (140 mg, 3.49 mmol) was added and thereaction mixture was stirred for 30 minutes at room temperature. Asolution of ethyl2-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)-2-propenoate(Intermediate 132, 830 mg) in dry dimethyl sulfoxide (5 mL) was addedand the reaction mixture was stirred for 30 minutes at room temperatureThe reaction was quenched with ice, diluted with brine (10 ml) and water(10 ml) and extracted with ethyl acetate (2×30 ml). The organic layerwas washed with water (3×15 ml) and brine (1×20 ml), dried over sodiumsulphate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 50 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 7:3 as eluent. Thisafforded the title compound as a colourless oil (780 mg).

¹H NMR (400 MHz, DMSO-d₆, 65° C.): δ ppm 7.25 (1H, t), 7.12 (1H, d),6.98 (1H, d), 5.19 (2H, s), 4.78 (2H, br.s), 4.01 (2H, q), 3.41 (3H, s),1.55-1.65 (2H, m), 1.13-1.21 (2H, m), 1.08 (3H, t), 0.94 (9H, s), 0.11(6H, s); UPLC_IPQC: 1.57 min, 395 [M+H]+.

Intermediate 134(2-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenyl)methanol

To a solution of ethyl1-(2-({[(1,1-dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-{[(methyloxy)methyl]oxy}phenyl)cyclopropanecarboxylate(Intermediate 133, 780 mg) in dry tetrahydrofuran (20 ml) at 0° C., asolution of LiAlH4 in THF (1M, 2.076 mL, 2.076 mmol) was slowly addedand the reaction mixture was stirred for 2 hours at the sametemperature. The reaction was quenched with water (10 ml) and brine (10ml) and diluted with ethyl acetate (30 ml). The solid was filtered offand two phases were separated. The aqueous layer was extracted withethyl acetate (30 ml) and the combined organic layers were dried oversodium sulphate, filtered and evaporated to afford the alcoholintermediate as a colourless oil. It was dissolved in drytetrahydrofuran (20.00 mL) and TBAF 1M solution in THF (2.076 mL, 2.076mmol) was slowly added at 0° C. The reaction mixture was stirred for 1hour at the same temperature. The reaction was quenched with water (10ml) and brine (10 ml) and extracted with ethyl acetate (2×30 ml). Theorganic layer was dried over sodium sulphate, filtered and evaporatedand the residue was purified by flash chromatography (Biotage system) onsilica gel using a 50 g SNAP column and cyclohexane/ethyl acetate aseluents from 7:3 to 3:7. This afforded the title compound (450 mg) as awhite crystal solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.16 (1H, t), 7.07 (1H, d), 6.92 (1H,d), 5.17-5.24 (2H, m), 5.00 (1H, t), 4.67-4.74 (3H, m), 3.65-3.76 (1H,m), 3.41 (3H, s), 3.11-3.21 (1H, m), 0.83-0.94 (2H, m), 0.65-0.77 (1H,m), 0.51-0.61 (1H, m).

Intermediate 1355-{[(methyloxy)methyl]oxy}-1H-spiro[2-benzopyran-4,1′-cyclopropane]

To a solution of(2-[1-(hydroxymethyl)cyclopropyl]-3-{[(methyloxy)methyl]oxy}phenyl)methanol(Intermediate 134, 450 mg) in dry tetrahydrofuran (10 ml) at 0° C., asolution of BuLi in hexane (1.6M, 1.180 mL, 1.889 mmol) was slowly addedand the reaction mixture was stirred for 15 minutes at the sametemperature. A solution of tosyl chloride (360 mg, 1.889 mmol) in drytetrahydrofuran (5 ml) was slowly added and the reaction mixture wasstirred for 15 minutes at 0° C. A second equivalent of a solution ofBuLi in hexane (1.6M, 1.180 mL, 1.889 mmol) was added and the reactionmixture was stirred for 30 minutes at the same temperature. The reactionwas quenched with an aqueous saturated solution of NaHCO₃ (10 ml)diluted with water (10 ml) and extracted with ethyl acetate (2×30 ml).The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 25 g SNAP column and cyclohexane/ethylacetate as eluents from 100:0 to 8:2. This afforded the title compound5-{[(methyloxy)methyl]oxy}-1H-spiro[2-benzopyran-4,1′-cyclopropane] as acolourless oil (385 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.04 (1H, t), 6.83 (1H, d), 6.68 (1H,d), 5.11 (2H, s), 4.75 (2H, s), 3.49 (2H, s), 3.37 (3H, s), 1.64-1.70(2H, m), 0.56-0.61 (2H, m).

Intermediate 136 1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-ol

To a solution of5-{[(methyloxy)methyl]oxy}-1H-spiro[2-benzopyran-4,1′-cyclopropane](Intermediate 135, 380 mg) in methanol (10 mL) HCl 10% in water (1.048mL, 3.45 mmol) was added and the reaction mixture was stirred overnightat 50° C. The combined solvents were evaporated and the residue waspurified by flash chromatography (Biotage system) on silica gel using a25 g SNAP column and cyclohexane/ethyl acetate as eluents from 100:0 to8:2. This afforded the title compound as a white solid (260 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.18 (1H, s), 6.88 (1H, t), 6.56 (1H,d), 6.47 (1H, d), 4.71 (2H, s), 3.47 (2H, s), 1.69-1.74 (2H, m),0.47-0.52 (2H, m).

Intermediate 1375-nitro-2-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)pyridine

To a solution of 1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-ol(Intermediate 136, 150 mg) in dry N,N-dimethylformamide (4 ml) potassiumcarbonate (176 mg, 1.277 mmol) and then 2-chloro-5-nitropyridine (148mg, 0.936 mmol) were added and the reaction mixture was stirred for 1 hat 110° C. After cooling the reaction was quenched with brine (1 ml),diluted with water (5 ml) and extracted with ethyl acetate (3×15 ml).The organic layer was washed with ice cold brine (2×10 ml), dried oversodium sulphate, filtered and evaporated and the residue was purified byflash chromatography (Biotage system) on silica gel using a 25 g SNAPcolumn and cyclohexane/ethyl acetate as eluents from 100:0 to 7:3. Thisafforded the title compound as a colourless gum (240 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.05 (1H, d), 8.64 (1H, dd), 7.24 (1H,d), 7.20 (1H, t), 7.01 (1H, d), 6.88 (1H, d), 4.86 (2H, s), 3.53 (2H,s), 1.23-1.27 (2H, m), 0.63-0.68 (2H, m); UPLC_IPQC: 1.08 min, 299[M+H]+.

Intermediate 138641H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinamine

To a solution of5-nitro-2-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)pyridine(Intermediate 137, 238 mg) in tetrahydrofuran (10 mL)/water (5 mL) iron(223 mg, 3.99 mmol) and then ammonium chloride (213 mg, 3.99 mmol) wereadded and the reaction mixture was stirred overnight at roomtemperature. The catalyst was filtered off and the solution was dilutedwith an aqueous saturated solution of NaHCO₃ and extracted with ethylacetate (2×30 ml). The organic layer was dried over sodium sulphatefiltered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 25 g SNAP columnand cyclohexane/ethyl acetate as eluents from 7:3 to 3:7. This affordedthe title compound (180 mg) as a light yellow solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.51 (1H, d), 7.07 (1H, dd), 7.04 (1H,t), 6.80 (1H, d), 6.69 (1H, d), 6.53 (1H, d), 5.06 (2H, br.s), 4.81 (2H,s), 3.53 (2H, s), 1.49-1.53 (2H, m), 0.58-0.62 (2H, m).

Intermediate 139

S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline (100mg, 0.460 mmol) in dry tetrahydrofuran (THF) (10 ml)2,2′-dithiodipyridine (254 mg, 1.151 mmol) and triphenylphosphine (302mg, 1.151 mmol) were added. The mixture was stirred at room temperaturefor 3 hours. THF was evaporated under vacuum. The residue was purifiedby flash chromatography on silica gel using a 25 g the title compound(78 mg, 0.251 mmol) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.59 (1H, d), 7.87 (1H, t), 7.70 (1H,br.s.), 7.51 (1H, d), 7.41 (t, 1H), 2.02-1.84 (1H, m), 1.74-1.60 (1H,m), 1.43 (9H, s), 1.33 (3H, s), 0.81 (3H, t).

UPLC_IPQC: 1.01 min, 311 [M+H]+.

Intermediate 1401,1-dimethylethyl[(1R)-1-methyl-1-({[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate

To a solution of6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinamine(Intermediate 138, mg, 0.242 mmol) in dry toluene (4 mL) S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 75 mg, 0.242 mmol) was added and the reaction mixturewas stirred for 3 hours at 150° C. After cooling the solvent was removedunder reduced pressure and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 9:1 to cyclohexane/ethyl acetate 1:1 aseluent affording the title compound as a white solid (64 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.63 (1H, br.s), 8.30 (1H, br.s),8.01-8.16 (1H, m), 7.11 (1H, t), 6.87-6.95 (3H, m), 6.69 (1H, d), 4.83(2H, s), 3.53 (2H, s), 1.70-1.90 (2H, m), 1.26-1.47 (14H, m), 0.78 (3H,t), 0.58-0.65 (2H, m); UPLC_IPQC: 1.14 min, 468 [M+H]+.

Intermediate 141N¹-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-D-isovalinamide

To a solution of1,1-dimethylethyl[(1R)-1-methyl-1-({[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]amino}carbonyl)propyl]carbamate(Intermediate 140, 62 mg) in dry dichloromethane (6 mL) at 0° C. TFA (2ml, 26.0 mmol) was slowly added and the reaction mixture was stirred for3 hours at the same temperature. The reaction mixture was diluted withdichloromethane (15 ml) and an aqueous saturated solution of NaHCO₃ wasadded while the pH was allowed to reach ˜8. Two phases were separatedand the organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 10 g SNAP column anddichloromethane/methanol as eluents from 99:1 to 95:5. This afforded thetitle compound as a white solid (42 mg, 86% yield).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.40 (1H, d), 8.16 (1H, dd), 7.11 (1H,t), 0.94 (1H, d), 6.90 (1H, d), 6.69 (1H, d), 4.84 (2H, s), 3.53 (2H,s), 1.66-1.78 (1H, m), 1.46-1.58 (1H, m), 1.37-1.42 (2H, m), 1.24 (3H,s), 0.83 (3H, t), 0.59-0.64 (2H, m); UPLC_IPQC: 0.69 min, 368 [M+H]+.

Intermediate 1421,1-dimethylethyl[2-({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(59.2 mg, 0.292 mmol) DIPEA (0.068 mL, 0.389 mmol) and then HATU (111mg, 0.292 mmol) were added and the reaction mixture was stirred for 15minutes at r.t.2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine(Intermediate 65, 50 mg, 0.194 mmol) was then added and the reactionmixture was stirred for 18 hours at 50° C. The reaction was quenchedwith brine (1 ml), diluted with water (2 ml) and extracted with ethylacetate (2×5 ml). The organic layer was dried (Na₂SO₄), filtered andevaporated and The residue was purified by flash chromatography (Biotagesystem) on silica gel using a column SNAP 25 g and cyclohexane tocyclohexane/ethyl acetate 7:3 as eluents affording the title compound(31 mg) as a white solid.

UPLC: 0.75 min, 443 [M+H]+.

Intermediate 143N1-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-2-methylalaninamide

To a solution of1,1-dimethylethyl[2-({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 142, 30 mg, 0.068 mmol) in dry Dichloromethane (DCM) (1.6mL) at 0° C. TFA (0.4 ml, 5.19 mmol) was slowly added and the reactionmixture was stirred for 1 h at the same temperature. The solvent and theexcess of TFA were evaporated end the residue was dissolved in DCM (5ml) and an aqueous saturated solution of NaHCO₃ was added while the pHwas allowed to reach ˜8-9. Two phases were separated and the organiclayer was dried (Na₂SO₄), filtered and evaporated affording the titlecompound (20 mg) as a white solid.

UPLC: 0.51 min, 343 [M+H]+.

Intermediate 1441,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]-2-methylpropyl}carbamate

To a solution of Boc-D-Valine (63.3 mg, 0.292 mmol) inN,N-Dimethylformamide (DMF) (2 mL) DIPEA (0.068 mL, 0.389 mmol) and thenHATU (111 mg, 0.292 mmol) were added and the reaction mixture wasstirred for 15 minutes at r.t.2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine(Intermediate 65, 50 mg, 0.194 mmol) was then added and the reactionmixture was stirred for 5 hours at 50° C. The reaction was quenched withbrine (1 ml), diluted with water (2 ml) and extracted with ethyl acetate(2×5 ml). The organic layer was dried (Na₂SO₄), filtered and evaporatedand The residue was purified by flash chromatography (Biotage system) onsilica gel using a column SNAP 25 g and cyclohexane to cyclohexane/ethylacetate 7:3 as eluent affording the title compound1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]-2-ethylpropyl}carbamate(55 mg, 0.120 mmol, 62.0% yield) as a white solid.

Intermediate 145(2R)-2-amino-N-[2-[(3,3-dimethyl-2H-benzofuran-4-yl)oxy]pyrimidin-5-yl]-3-methyl-butanamide

To a solution of 1,1-dimethylethyl{(1R)-1-[({2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]-2-methylpropyl}carbamate(Intermediate 144, 52 mg, 0.114 mmol) in dry Dichloromethane (DCM) (2mL) at 0° C. TFA (0.5 ml, 6.49 mmol) was slowly added and the reactionmixture was stirred for 1 h at the same temperature. The solvent and theexcess of TFA were evaporated end the residue was dissolved in DCM (5ml) and an aqueous saturated solution of NaHCO₃ was added while the pHwas allowed to reach ˜8-9. Two phases were separated and the organiclayer was dried (Na₂SO₄), filtered and evaporated affording the titlecompound (40 mg) as a white solid.

UPLC: 0.53 min, 357 [M+H]+.

Intermediate 146 tert-butylN-[(1R)-1-methyl-1-[[6-(3-methylchroman-5-yl)oxy-3-pyridyl]carbamoyl]propyl]carbamate

In a 8 mL 6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinamine(Intermediate 119, 82.4 mg, 0.0305 mmol),(2R)-2-(tert-butoxycarbonylamino)-2-methyl-butanoic acid (59.7 mg, 0.275mmol) and DIPEA (0.080 mL, 0.458 mmol) were dissolved inN,N-Dimethylformamide (DMF) (2 mL) to give a pale yellow solution. HATU(151 mg, 0.397 mmol) was added. The reaction mixture was stirred at roomtemperature over week-end. The reaction mixture was evaporated in vacuoand the residue was purified by flash chromatography on silica gel usinga column SNAP 25 g and cyclohexane/ethyl acetate from 3:1 to 1:2 aseluents affording the title compound (55.8 mg) as white solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.08-8.16 (2H, m), 7.10 (1H, t), 6.84 (1H,d), 6.70 (1H, d), 6.59 (1H, d), 6.13 (1H, br. s.), 4.90 (1H, br. s.),4.12-4.22 (1H, m), 3.59-3.75 (1H, m), 2.59-2.92 (1H, m), 1.85-2.25 (3H,m), 1.69-1.84 (1H, m), 1.51 (3H, s), 1.45 (9H, s), 0.91-1.05 (6H, m).UPLC_ipqc: 1.22 min, 456 [M+H]+.

Intermediate 147(2R)-2-amino-2-methyl-N-[6-(3-methylchroman-5-yl)oxy-3-pyridyl]butanamide

In a 50 mL round-bottomed flask tert-butylN-[(1R)-1-methyl-1-[[6-(3-methylchroman-5-yl)oxy-3-pyridyl]carbamoyl]propyl]carbamate(Intermediate 146, 55.8 mg, 0.104 mmol) was dissolved in Dichloromethane(3 mL) to give a pale yellow solution. The reaction mixture was cooledat 0° C. and TFA (2 mL, 26.0 mmol) was added. The reaction mixture wasstirred at 0° C. for 2 hours. The reaction mixture was evaporated invacuo to give the crude product as a yellow oil. The sample was chargedon a 2 g SCX cartridge. It was then flushed with 36 mL of MeOH followedby 25 mL of 2M solution of ammonia in MeOH. The ammonia eluate wasevaporated in vacuo affording the title compound (32.9 mg) as paleyellow oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 9.88 (1H, br. s.), 8.14-8.28 (2H, m), 7.10(1H, t), 6.83 (1H, d), 6.70 (1H, dd), 6.59 (1H, dd), 4.12-4.21 (1H, m),3.61-3.72 (1H, m), 2.72-2.84 (1H, m), 2.04-2.24 (2H, m), 1.92-2.04 (1H,m), 1.52-1.84 (6H, m), 1.00 (3H, d), 0.94 (3H, t). UPLC_ipqc: 0.76 min,356 [M+H]+.

Intermediate 1481,1-dimethylethyl[2-({6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate

In a 8 mL vial6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 108, 55 mg, 0.215 mmol) was dissolved inN,N-Dimethylformamide (2 mL) to give a colourless solution. DIPEA (0.056mL, 0.322 mmol), N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine(52.3 mg, 0.258 mmol) and, TBTU (83 mg, 0.258 mmol) were added. Thereaction mixture was shake N at 60° C. overnight., Additional DIPEA (0.1mL), N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (105 mg) andTBTU (170 mg) were added. The reaction mixture was shaken at 60° C. foradditional 10 hours. The reaction mixture was evaporated in vacuo andthe residue was purified by flash chromatography on silica gel using acolumn SNAP 25 g and cyclohexane/ethyl acetate from 2:1 to 1:1 aseluents affording the title compound (36.5 mg) as colorless oil solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.12-8.17 (2H, m), 7.30 (1H, t), 7.05 (1H,d), 6.95 (1H, d), 6.89 (1H, d), 6.16 (1H, br. s.), 5.20-5.27 (1H, m),5.17 (1H, dd), 5.10 (1H, d), 4.92 (1H, br. s.), 1.84-1.97 (1H, m),1.66-1.78 (1H, m), 1.63 (6H, s), 1.45 (9H, s), 0.88 (3H, t). UPLC_B:0.89 min, 442 [M+H]+.

Intermediate 149N1-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2-methylalaninamide

In a 50 mL round-bottomed flask1,1-dimethylethyl[2-({6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}amino)-1,1-dimethyl-2-oxoethyl]carbamate(Intermediate 148, 36.5 mg, 0.066 mmol) was dissolved in Dichloromethane(3 mL) to give a colourless solution. The reaction mixture was cooled at0° C. and TFA (1.5 mL, 19.47 mmol) was added at that temperature. Thereaction mixture was stirred at 0° C. for 1.5 hours. The reactionmixture was evaporated in vacuo affording and the residue was charged ona 2 g SCX cartridge. It was then flushed with 15 mL of MeOH followed by15 mL of 2M solution of ammonia in MeOH. The ammonia eluate wasevaporated in vacuo affording the title compound (17.2 mg) as colorlessoil.

¹H NMR (400 MHz, CDCl₃) δ ppm 9.91 (1H, br. s.), 8.26 (1H, dd), 8.20(1H, d), 7.30 (1H, t), 7.05 (1H, d), 6.94 (1H, d), 6.90 (1H, d),5.21-5.27 (1H, m), 5.17 (1H, dd), 5.10 (1H, d), 1.83-1.98 (1H, m),1.68-1.79 (1H, m), 1.47 (6H, s), 0.88 (3H, t). UPLC_B: 0.74 min, 342[M+H]+.

Intermediate 150 2,4-bis(methoxymethoxy)-1-methyl-benzene

To a solution of 4-methylbenzene-1,3-diol (4 g, 32.26 mmol) in dryN,N-Dimethylformamide (30 ml) at 0° C. sodium hydride (60% dispersion inmineral oil) (3.87 g, 96.78 mmol) was added and the reaction mixture wasstirred for 15 minutes at the same temperature. MOM-Cl (7.35 ml, 96.78mmol) was quickly added and the reaction mixture was stirred for 1 hourwhile the temperature was allowed to reach room temperature. Thereaction was quenched with brine (40 ml) and extracted with ethylacetate (3×80 ml). The organic layer was washed with ice cold brine(2×50 ml), dried over sodium sulphate, filtered and evaporated and theresidue was purified by flash chromatography (Biotage system) on silicagel using a 100 g SNAP column and cyclohexane to cyclohexane/ethylacetate 8:2 as eluents affording the title compound (6.1 g) as acolourless oil.

LC/MS: QC_(—)3_MIN: Rt=1.811 min; 213 [M+H]+.

Intermediate 151 ethyl2-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]-2-oxo-acetate

To a solution of 2,4-bis(methoxymethoxy)-1-methyl-benzene (Intermediate150, 5.5 g, 25.94 mmol) in dry tetrahydrofuran (50 ml) at roomtemperature BuLi 1.6M in hexane (19.45 ml, 31.13 mmol) was added and thereaction mixture was stirred for 30 minutes at the same temperature. Themixture was cooled to −78° C. and it was added (via cannulation) to asolution of ethyl chlorooxoacetate (4.35 ml, 38.9 mmol) in drytetrahydrofuran (30 ml) at −78° C. The reaction mixture was stirred at−78° C. for 30 minutes. The reaction was quenched with water (20 ml),diluted with brine (50 ml) and extracted with ethyl acetate (2×100 ml).Combined organic layers were dried over sodium sulphate, filtered andevaporated. The residue was purified by flash chromatography (Biotagesystem) on silica gel using a 100 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 8:2 as eluent affording the title compound(4.65 g) as a light yellow oil.

LC/MS: QC_(—)3_MIN: Rt=1865 min.

Intermediate 152 ethyl2-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]prop-2-enoate

To a suspension of methyltriphenylphosphonium bromide (8.78 g, 24.6mmol) in dry tetrahydrofuran (50 ml) at 0° C. KHMDS 0.5M solution intoluene (44.22 ml, 22.11 mmol) was slowly added and the reaction mixturewas stirred for 15 minutes at 0° C. and for 45 minutes at roomtemperature. The reaction mixture was cooled to 0° C. and it was slowlyadded to a solution of ethyl2-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]-2-oxo-acetate (Intermediate151, 4.6 g, 14.74 mmol) in dry tetrahydrofuran (25 mL) at 0° C. and thereaction mixture was stirred for 2 hours at 0° C. The reaction wasquenched with water (50 ml), diluted with brine (50 ml) and extractedwith ethyl acetate (2×100 ml). The organic layer was dried over sodiumsulphate, filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 100 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 8:2 as eluents affordingthe title compound (3.8 g) as a colourless oil.

LC/MS: QC_(—)3_MIN: Rt=1.930 min.

Intermediate 153 ethyl1-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]cyclopropanecarboxylate

To a solution of trimethylsulfoxonium iodide (4.4 g, 20 mmol) in drydimethyl sulfoxide (30 mL) sodium hydride (60% dispersion in mineraloil) (0.720 g, 18 mmol) was added and the reaction mixture was stirredfor 1 hour at room temperature. A solution of ethyl2-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]prop-2-enoate (Intermediate152, 3.5 g, 11.29 mmol) in dry dimethyl sulfoxide (15 mL) was slowlyadded and the reaction mixture was stirred for 1 hour at roomtemperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (10 ml), diluted with water (40 ml) andextracted with ethyl acetate (2×100 ml). The organic layer was washedwith water (2×50 ml), dried over sodium sulphate, filtered andevaporated. The residue was purified by flash chromatography (Biotagesystem) on silica gel using a 100 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 8:2 as eluents affording the title compound(3.1 g) as a colourless oil.

LC/MS: QC_(—)3_MIN: Rt=2.028 min.

Intermediate 1542-[1-(hydroxymethyl)cyclopropyl]-3-(methoxymethoxy)-6-methyl-phenol

To a solution of ethyl1-[2,6-bis(methoxymethoxy)-3-methyl-phenyl]cyclopropanecarboxylate(Intermediate 153, 300 mg, 0.93 mmol) in ethanol (10 ml) HCl 6N in water(0.4 mL, 2.4 mmol) was added and the reaction mixture was stirredovernight at 50° C. Combined solvents were removed under reducedpressure. The residue was suspended in dry toluene (10 mL) and thesolvent evaporated. The obtained residue was dissolved in drytetrahydrofuran (10 ml), the mixture was cooled to 0° C. and NaH (60%dispersion in mineral oil) (80 mg, 2 mmol) was added and the reactionmixture was stirred for 30 minutes at the same temperature. MOM-Cl(0.083 mL, 1.1 mmol) was then added and the reaction mixture was stirredfor 1 hour at 0° C. LiAlH₄ (1M in THF, 1.2 ml, 1.2 mmol) was added andthe reaction mixture was further stirred for 1 hour at the sametemperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (10 ml), diluted with water (20 ml) andextracted with ethyl acetate (2×50 ml). Combined organic layers weredried over sodium sulphate, filtered and evaporated and the residue waspurified by flash chromatography (Biotage system) on silica gel using a25 g SNAP column and cyclohexane to cyclohexane/ethyl acetate 7:3 aseluents affording the title compound (70 mg) as a white solid.

LC/MS: QC_(—)3_MIN: Rt=1.690 min; 239 [M+H]+.

Intermediate 1554-(methoxymethoxy)-7-methyl-spiro[2H-benzofuran-3,1′-cyclopropane]

To a solution of2-[1-(hydroxymethyl)cyclopropyl]-3-(methoxymethoxy)-6-methyl-phenol(Intermediate 154, 65 mg, 0.27 mmol) in dry tetrahydrofuran (5 ml)triphenylphosphine (84 mg, 0.32 mmol) was added and the reaction mixturewas stirred until complete dissolution of PPh3. DIAD (0.056 ml, 0.285mmol) was then added dropwise and the reaction mixture was stirred for30 minutes at room temperature. The solvent was removed under reducedpressure and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 10 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 8:2 as eluents affording the title compound(40 mg) as a light yellow oil.

LC/MS: QC_(—)3_MIN: Rt=2.024 min; 221 [M+H]+.

Intermediate 156 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol

To a solution of4-(methoxymethoxy)-7-methyl-spiro[2H-benzofuran-3,1′-cyclopropane](Intermediate 155, 38 mg, 0.17 mmol) in ethanol (5 ml), HCl 6N in water(0.1 mL, 0.6 mmol) was added and the reaction mixture was stirred for 4days at room temperature. Combined solvents were removed under reducedpressure and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 10 g SNAP column and cyclohexane tocyclohexane/ethyl acetate 7:3 as eluents affording the title compound(24 mg) as a light orange solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 9.02 (1H, s), 6.65 (1H, d), 6.06 (1H,d), 4.36 (2H, s), 2.02 (3H, s), 1.40-1.44 (2H, m), 0.77-0.82 (2H, m).ROESY (400 MHz, DMSO-d₆): NOE correlation between proton at 6.65 ppm andprotons (CH3) at 2.02 ppm, NOE correlation between proton at 9.02 ppmand proton at 6.06 ppm. LC/MS: QC_(—)3_MIN: Rt=1.647 min; 177 [M+H]+.

Intermediate 1572-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-5-nitro-pyridine

To a solution of 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 156, 176 mg, 1 mmol) in dry DMF (4 ml) potassium carbonate(207 mg, 1.5 mmol) and then 2-chloro-5-nitropyridine (158 mg, 1 mmol)were added and the reaction mixture was stirred for 2 hours at 80° C.After cooling the reaction mixture was quenched with water (2 ml),diluted with brine (10 ml) and extracted with ethyl acetate (2×20 ml).The organic layer was dried over sodium sulfate, filtered and evaporatedaffording the title compound (270 mg) as an orange solid that was usedin the next step as crude material without further purification.

LC/MS: QC_(—)3_MIN: Rt=2.138 min; 299 [M+H]+.

Intermediate 1586-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyridin-3-amine

To a solution of2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-5-nitro-pyridine(Intermediate 157, 265 mg) in tetrahydrofuran (5 ml)/water (2.5 ml) iron(245 mg, 4.45 mmol) and then ammonium chloride (238 mg, 4.45 mmol) wereadded and the reaction mixture was stirred overnight at roomtemperature. The catalyst was filtered off and the residue was dilutedwith an aqueous saturated solution of NaHCO₃ (5 ml) and extracted withethyl acetate (3×10 ml). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 1:1 aseluents affording the title compound (203 mg) as a light yellow solid.

LC/MS: QC_(—)3_MIN: Rt=1.740 min; 269 [M+H]+.

Intermediate 159 tert-butylN-[(1R)-1-[[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]carbamoyl]propyl]carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (36 mg,0.18 mmol) in dry DMF (1 ml) DIPEA (52 μl, 0.3 mmol) and then HATU (65mg, 0.17 mmol) were added and the reaction mixture was stirred for 15minutes at r.t.6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyridin-3-amine(Intermediate 158, 40 mg, 0.15 mmol) was then added and the reactionmixture was stirred for 4 hours at room temperature. The reaction wasquenched with water (2 ml) diluted with brine (5 ml) and extracted withethyl acetate (2×10 ml). The organic layer was dried (Na₂SO₄), filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 10 g SNAP column andcyclohexane/ethyl acetate 90:10 to cyclohexane/ethyl acetate 60:40 aseluents affording the title compound (57 mg) as a white solid.

LC/MS: QC_(—)3_MIN: Rt=2.190 min; 454 [M+H]+.

Intermediate 160(2R)-2-amino-N-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]butanamide

To a solution of tert-butylN-[(1R)-1-[[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]carbamoyl]propyl]carbamate(Intermediate 159, 55 mg) in dry DCM (3 ml) at 0° C. TFA (1 ml) wasslowly added and the reaction mixture was stirred for 3 hours at thesame temperature. The solvent and the excess of TFA were removed underreduced pressure and the residue was diluted with DCM (10 ml) and anaqueous saturated solution NaHCO₃ was added while the pH was allowed toreach ˜8. Two phases were separated and the organic layer was dried(Na₂SO₄), filtered and evaporated affording the title compound (41 mg)as white solid.

LC/MS: QC_(—)3_MIN: Rt=1.792 min; 354 [M+H]+.

Intermediate 1612-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-5-nitro-pyrimidine

To a solution of 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 156, 176 mg, 1 mmol) in dry Acetonitrile (4 ml) potassiumcarbonate (207 mg, 1.5 mmol) and then 2-chloro-5-nitropyrimidine (159mg, 1 mmol) were added and the reaction mixture was stirred for 24 hoursat 80° C. After cooling the reaction mixture was quenched with water (2ml), diluted with brine (10 ml) and extracted with ethyl acetate (2×20ml). The organic layer was dried over sodium sulfate, filtered andevaporated affording the title compound (258 mg) as an orange solid thatwas used in the next step as crude material without furtherpurification.

LC/MS: QC_(—)3_MIN: Rt=2.007 min; 300 [M+H]+.

Intermediate 1622-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-amine

To a solution of2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-5-nitro-pyrimidine(Intermediate 161, 255 mg) in tetrahydrofuran (5 ml)/water (2.5 ml) iron(234 mg, 4.25 mmol) and then ammonium chloride (227 mg, 4.25 mmol) wereadded and the reaction mixture was stirred for 48 hours at roomtemperature. The catalyst was filtered off and the residue was dilutedwith an aqueous saturated solution of NaHCO₃ (5 ml) and extracted withethyl acetate (3×10 ml). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 4:6 aseluents affording the title compound (52 mg) as a light orange solid.

LC/MS: QC_(—)3_MIN: Rt=1.746 min; 270 [M+H]+.

Intermediate 163 tert-butylN-[(1R)-1-[[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]carbamoyl]propyl]carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (45 mg,0.222 mmol) in dry DMF (1 ml) DIPEA (870, 0.5 mmol) and then HATU (80mg, 0.21 mmol) were added and the reaction mixture was stirred for 15minutes at r.t.2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-amine(Intermediate 162, 50 mg, 0.185 mmol) was then added and the reactionmixture was stirred for 6 hours at room temperature. The reaction wasquenched with water (2 ml) diluted with brine (5 ml) and extracted withethyl acetate (2×10 ml). The organic layer was dried (Na₂SO₄), filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 10 g SNAP column andcyclohexane/ethyl acetate 90:10 to cyclohexane/ethyl acetate 60:40 aseluents affording the title compound (45 mg) as a white solid.

LC/MS: QC_(—)3_MIN: Rt=2.109 min; 455 [M+H]+.

Intermediate 164(2R)-2-amino-N-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]butanamide

To a solution of tert-butylN-[(1R)-1-[[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]carbamoyl]propyl]carbamate(Intermediate 163, 42 mg) in dry DCM (3 ml) at 0° C. TFA (1 ml) wasslowly added and the reaction mixture was stirred for 3 hours at thesame temperature. The solvent and the excess of TFA were removed underreduced pressure and the residue was diluted with DCM (10 ml) and anaqueous saturated solution NaHCO₃ was added while the pH was allowed toreach ˜8. Two phases were separated and the organic layer was dried(Na₂SO₄), filtered and evaporated affording the title compound (25 mg)as light yellow gum.

LC/MS: QC_(—)3_MIN: Rt=1.688 min; 355 [M+H]+.

Intermediate 165(5R)-3-(2-chloropyrimidin-5-yl)-5-ethyl-5-methyl-imidazolidine-2,4-dione

To a solution of triphosgene (1.38 g, 4.65 mmol) in Ethyl acetate (20ml) at 0° C. a solution of 2-chloro-5-aminopyrimidine (1 g, 7.75mmol)/DIPEA (8 ml, 4.65 mmol) in ethyl acetate (40 ml) was slowly added(20 minutes) and the reaction mixture was stirred for 15 minutes at thesame temperature. Maintaining the reaction mixture at 0° C., vacuum wasapplied (10 minutes) for removing the excess of phosgene. A solution ofDMAP (0.945 g, 7.75 mmol) in ethyl acetate/dichloromethane 1:1 (8 ml)was added and the reaction mixture was stirred for 5 minutes at the sametemperature. A solution of methyl (R)-2-amino-2-methyl-butyratehydrochloride (2.59 g, 15.5 mmol) in ethyl acetate (30 ml) was slowlyadded (15 minutes) at 0° C. and the reaction mixture was stirred for 30minutes at the same temperature. The reaction was quenched with aqueousbuffer (pH3) while the pH was allowed to reach ˜5-6 and two phases wereseparated. The organic layer was washed with aqueous buffer (pH3) (2×20ml) and then brine (20 ml), dried (Na₂SO₄), filtered and evaporatedaffording the urea intermediate as orange foam.

The urea was dissolved in MeOH (20 ml), NaOMe (0.41 g, 7.75 mmol) wasadded and the reaction mixture was stirred for 15 minutes at r.t. Themixture was quenched with an aqueous saturated solution of ammoniumchloride (25 ml) and diluted with ethyl acetate (50 ml). Two phases wereseparated and the organic layer was washed with brine (2×20 ml), dried(Na₂SO₄), filtered and evaporated. The residue was triturated with Et₂O(10 ml) and the solid collected affording the title compound (1.22 g) asa beige solid.

LC/MS: QC_(—)3_MIN: Rt=1.341 min; 255 [M+H]+.

Intermediate 1663-(2-chloropyrimidin-5-yl)-5,5-dimethyl-imidazolidine-2,4-dione

To a solution of triphosgene (1.38 g, 4.65 mmol) in Ethyl acetate (20ml) at 0° C. a solution of 2-chloro-5-aminopyrimidine (1 g, 7.75mmol)/DIPEA (8 ml, 4.65 mmol) in ethyl acetate (40 ml) was slowly added(20 minutes) and the reaction mixture was stirred for 15 minutes at thesame temperature. Maintaining the reaction mixture at 0° C., vacuum wasapplied (10 minutes) for removing the excess of phosgene. A solution ofDMAP (0.945 g, 7.75 mmol) in ethyl acetate/dichloromethane 1:1 (8 ml)was added and the reaction mixture was stirred for 5 minutes at the sametemperature. 2,2-Dimethylglycine methyl ester hydrochloride (2.37 g,15.5 mmol) in ethyl acetate (30 ml) was slowly added (15 minutes) at 0°C. and the reaction mixture was stirred for 30 minutes at the sametemperature. The reaction was quenched with aqueous buffer (pH3) whilethe pH was allowed to reach ˜5-6 and two phases were separated. Theorganic layer was washed with aqueous buffer (pH3) (2×20 ml) and thenbrine (20 ml), dried (Na₂SO₄), filtered and evaporated affording theurea intermediate as orange foam.

The urea was dissolved in MeOH (20 ml), NaOMe (0.41 g, 7.75 mmol) wasadded and the reaction mixture was stirred for 15 minutes at r.t. Themixture was quenched with an aqueous saturated solution of ammoniumchloride (25 ml) and diluted with ethyl acetate (50 ml). Two phases wereseparated and the organic layer was washed with brine (2×20 ml), dried(Na₂SO₄), filtered and evaporated. The residue was triturated with Et₂O(10 ml) and the solid collected affording the title compound (1.08 g) asan orange solid.

LC/MS: QC_(—)3_MIN: Rt=1.062 min; 241 [M+H]+.

Intermediate 167 methyl 3-(2-methylallyloxy)benzoate

Methyl 3-hydroxybenzoate (1 g, 6.57 mmol) was dissolved in DMF (10 ml)to give a colorless solution. To the solution potassium carbonate (1.089g, 7.88 mmol) and 3-Bromo-2-methylpropene (0.729 ml, 7.23 mmol) wereadded. The reaction mixture was heated to 90° C. and stirred for 1 h.After cooling the reaction mixture was diluted with water and extractedwith ethyl acetate. The organic phase was washed with water, dried overNa₂SO₄ and evaporated under vacuum to afford the title compound (1.180mg)

LC/MS: QC_(—)3_MIN: Rt=2.073 min; 207 [M+H]+.

Intermediate 168 methyl 3-hydroxy-2-(2-methylallyl)benzoate

Methyl 3-(2-methylallyloxy)benzoate (Intermediate 167, 1.100 g, 5.3mmol) was dissolved in 1-Methyl-2-pyrrolidinone (12 ml) and heated to200° C. The solution was stirred for 30 h at the same temperature. Aftercooling, the mixture was diluted with water and extracted with ethylacetate. Organic phase was dried over Na₂SO₄ and evaporated in vacuo toafford a crude product that was purified via Biotage SP1 withCyclohexane/EtOAc as eluents (from 10/0 to 7/3 for 12 CV, 50 g SNAPSilica column). Fractions were collected and evaporated to afford thetitle compound (507 mg).

LC/MS: QC_(—)3_MIN: Rt=1.772 min; 207 [M+H]+.

Intermediate 169 3-(hydroxymethyl)-2-(2-methylallyl)phenol

Methyl 3-hydroxy-2-(2-methylallyl)benzoate (Intermediate 168, 410 mg,1.99 mmol) was dissolved in tetrahydrofuran (5 ml) to give a colourlesssolution. The reaction mixture was cooled at 0° C. A solution of LiAlH₄2M in THF (1.09 ml, 2.19 mmol) was added dropwise and the reactionmixture was stirred at 0° C. for 30 min. After this time the reactionmixture was poured into ice and diluted with 60 ml of ethyl acetate.Phases were separated, the organic phase was dried over Na₂SO₄ andevaporated under vacuum to afford the title compound (360 mg) as acolorless oil.

LC/MS: QC_(—)3_MIN: Rt=1.192 min.

Intermediate 170 3,3-dimethylisochroman-5-ol

3-(hydroxymethyl)-2-(2-methylallyl)phenol (Intermediate 169, 360 mg, 2mmol) was dissolved in ethyl acetate (20 ml), two drops of sulfuric acidwere added to the solution that was stirred for 4 hours at roomtemperature. After this time the reaction was diluted with water (40 ml)and ethyl acetate (40 ml). Phases were separated and the organic layerwas dried over Na₂SO₄ and evaporated in vacuo to afford a colourlessoil. The oil was triturated with cyclohexane to obtain a white solidthat was filtered, washed with cyclohexane (20 ml) and dried in vacuo toafford the title compound (130 mg).

LC/MS: QC_(—)3_MIN: Rt=1.441 min.

Intermediate 171 2-(3,3-dimethylisochroman-5-yl)oxy-5-nitro-pyridine

To a solution of 3,3-dimethylisochroman-5-ol (Intermediate 170, 65 mg,0.36 mmol) in dry DMF (3 ml) potassium carbonate (207 mg, 1.5 mmol) andthen 2-chloro-5-nitropyridine (50.8 mg, 0.32 mmol) were added and thereaction mixture was stirred for 2 hours at 80° C. After cooling thereaction mixture was quenched with water (2 ml), diluted with brine (10ml) and extracted with ethyl acetate (2×20 ml). The organic layer wasdried over sodium sulfate, filtered and evaporated affording the titlecompound (80 mg) as an orange solid that was used in the next step ascrude without further purification.

LC/MS: QC_(—)3_MIN: Rt=2.027 min; 301 [M+H]+.

Intermediate 172 6-(3,3-dimethylisochroman-5-yl)oxypyridin-3-amine

To a solution of 2-(3,3-dimethylisochroman-5-yl)oxy-5-nitro-pyridine(Intermediate 171, 80 mg) in tetrahydrofuran (5 ml)/water (2.5 ml) iron(70 mg, 1.3 mmol) and then ammonium chloride (70 mg, 1.3 mmol) wereadded and the reaction mixture was stirred overnight at roomtemperature. The catalyst was filtered off and the residue was dilutedwith an aqueous saturated solution of NaHCO₃ (5 ml) and extracted withethyl acetate (3×10 ml). The organic layer was dried over sodiumsulfate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 4:6 aseluents affording the title compound (25 mg) as a white solid.

LC/MS: QC_(—)3_MIN: Rt=1.586 min; 271 [M+H]+.

Intermediate 173 [5-(methoxymethoxy)-2-methyl-phenyl]methanol

5-Hydroxy-2-methyl-benzoic acid (2 g, 13.3 mmol) was dissolved intetrahydrofuran (40 ml). The reaction mixture was cooled to 0° C. andsodium hydride (60% dispersion in mineral oil) (1.8 g, 39.5 mmol) wasadded portionwise. Chloro(methyloxy)methane (4 ml, 52 mmol) was added.The reaction mixture was stirred for 30 min at 0° C. The mixture waspoured into ice and extracted with ethyl acetate, two phases wereseparated and the organic layer was dried over Na₂SO₄ and evaporatedunder vacuum to give the crude product as a yellow oil. To thismaterial, dissolved in THF (20 ml), cooled at 0° C., LiAlH₄ 1M in THF(15 ml, 15 mmol) was added dropwise and the reaction mixture was stirredat 0° C. for 30 min. After this time the reaction mixture was pouredinto ice and diluted with 60 ml of ethyl acetate. Phases were separated,the organic phase was dried over Na₂SO₄ and evaporated under vacuum toafford a colorless oil that was purified by flash chromatography(Biotage system) on silica gel using a 100 g SNAP column and cyclohexaneto cyclohexane/ethyl acetate 75:25 as eluents affording the titlecompound (1.9 g) as a colorless oil.

LC/MS: QC_(—)3_MIN: Rt=1.351 min

Intermediate 174tert-butyl-[[5-(methoxymethoxy)-2-methyl-phenyl]methoxy]-dimethyl-silane

[5-(methoxymethoxy)-2-methyl-phenyl]methanol (Intermediate 173, 1.9 g,10 mmol) was dissolved in dichloromethane (10 ml) to give a colourlesssolution. 1H-imidazole (1.137 g, 16.7 mmol) andchloro(1,1-dimethylethyl)dimethylsilane (2.095 g, 13.9 mmol) were added.The reaction mixture immediately became a white suspension and wasstirred at room temperature for 30 minutes. The reaction mixture wasquenched with 10 ml of water and diluted with 10 ml of dichloromethane.Two phases were separated through a separating funnel. The organic phasewas dried over Na₂SO₄ and evaporated under vacuum and the residue waspurified by flash chromatography (Biotage system) on silica gel using a100 g SNAP column and cyclohexane to cyclohexane/ethyl acetate 50:50 aseluents affording the title compound (2.9 g) as a colorless oil.

LC/MS: QC_(—)3_MIN: Rt=2.437 min

Intermediate 1751-[2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-(methoxymethoxy)-3-methyl-phenyl]cyclobutanol

To a solution oftert-butyl-[[5-(methoxymethoxy)-2-methyl-phenyl]methoxy]-dimethyl-silane(Intermediate 174, 0.3 g, 1 mmol) in hexane (5 ml), BuLi (1.6M inhexane, 0.9 ml, 1.4 mmoli) was added at room temperature. The reactionmixture was stirred for 2 h and then added dropwise at −30° C. to asuspension of CeCl₃ (0.37 g, 1.5 mmoli) in dry THF (5 ml) that waspreviously stirred at room temperature overnight. After 45 min at −30°C., cyclobutanone (0.07 g, 1 mmoli) dissolved in THF (1 ml) was added.The reaction was stirred at the same temperature for and then quenchedwith ammonium chloride (20 ml) and extracted with Ethyl Acetate (2×20ml). Combined organic layers were dried over Na₂SO₄ and evaporated undervacuum to afford a colorless oil that was purified by flashchromatography (Biotage system) on silica gel using a 25 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 75:25 as eluents affordingthe title compound (0.05 g) as a colorless oil.

LC/MS: QC_(—)3_MIN: Rt=2.437 min

Intermediate 176 7-methylspiro[1H-isobenzofuran-3,1′-cyclobutane]-4-ol

To a solution of1-[2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-(methoxymethoxy)-3-methyl-phenyl]cyclobutanol(Intermediate 175, 0.05 g, 0.136 mmoli) in Ethyl Acetate (5 ml),Sulphuric Acid (96%, 2 drops) was added at room temperature and thereaction mixture was stirred for 2 h. Ethyl Acetate (20 ml) was addedand the organic phase was washed with brine (2×50 ml), dried over sodiumsulphate and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane to cyclohexane/ethyl acetate 7:3 as eluents affordingthe title compound (0.02 g) as a colourless oil.

LC/MS: QC_(—)3_MIN: Rt=1.720 min

Intermediate 177 Ethyl2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-(methoxymethoxy)benzoate

Under nitrogen flush, in a 2-necked 100 ml round-bottomed flask equippedwith a reflux condenser (flammed for 5 minutes under vacuum and then 3cycles of N2/vacuum),(1,1-dimethylethyl)(dimethyl){[(3-{[(methyloxy)methyl]oxy}phenyl)methyl]oxy}silane(Intermediate 103, 1.5 g, 5.31 mmol) was dissolved in hexane (20 ml) togive a colourless solution. Butyllithium 1.6N in hexane (4.31 ml, 6.9mmol) was added dropwise and the reaction mixture was stirred at roomtemperature. After 2 hours stirring in those conditions, the pale yellowreaction mixture was added to a solution of ethyl chloroformate (1.015ml, 10.62 mmol) in tetrahydrofuran at −78° C. After 30 min, the reactionwas quenched with a 2M aqueous solution of hydrochloric acid while thepH was allowed to reach ˜2 and diluted with 10 ml of ethyl acetate. Twophases were separated and the organic layer was dried over Na₂SO₄ andevaporated under vacuum affording the crude product as a yellowish oilthat was purified by flash chromatography (Biotage system) on silica gelusing cyclohexane to cyclohexane/ethyl acetate 8:2 as eluents affordingthe title compound (1.396 g) as a yellow pale oil.

LC/MS: QC_(—)3_MIN: Rt=2.413 min; 355 [M+H]+.

Intermediate 178 Ethyl2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-hydroxy-benzoate

Ethyl2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-(methoxymethoxy)benzoate(Intermediate 177, 950 mg, 2.68 mmol) was dissolved in dichloromethane(30 ml), the solution was cooled to 0° C. and trifluoroacetic acid (2ml) was added. After 3 h stirring at 0° C. water (20 ml) was added at 0°C. and two phases were separated. The organic layer was dried overNa₂SO₄ and evaporated in vacuo to afford a colourless oil that waspurified by flash chromatography (Biotage system) on silica gel usingcyclohexane to cyclohexane/ethyl acetate 7:3 as eluents affording thetitle compound (0.345 g) as a colorless oil.

LC/MS: QC_(—)3_MIN: Rt=2.463 min; 311 [M+H]+.

Intermediate 179 3,3-diethyl-1H-isobenzofuran-4-ol

To a 1M solution in THF of Ethylmagnesium bromide (10 ml, 5 mmol) in dryTHF (5 ml) at 0° C. a solution of ethyl2-[[tert-butyl(dimethyl)silyl]oxymethyl]-6-hydroxy-benzoate(Intermediate 178, 0.31 g, 1 mmol) in Et₂O (10 ml) was added in 15 min.The reaction mixture was stirred for 1.5 h at the same temperature andthen for additional 2 hours while the temperature was allowed to reachroom temperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (40 ml) and extracted with ethyl acetate(3×80 ml). The organic layer was washed with brine (2×50 ml), dried oversodium sulphate, filtered and evaporated. The residue was dissolved inTHF (5 ml) and tetrabutylammonium fluoride (1M in THF, 1.5 ml, 1.5 mmol)was added and the reaction mixture was stirred for 15 minutes. EthylAcetate (50 ml) was added and the combined organic layers were washedwith Ammonium Chloride (2×50 ml), dried over sodium sulphate, andconcentrated. The yellow solid obtained was triturated with EthylAcetate and Pentane (1:1, 5 ml) to give a white solid.

The solid was dissolved in Ethyl Acetate (5 ml) and Sulphuric Acid (96%,4 drops) was added at room temperature. The reaction was stirred for 2h, then Ethyl Acetate (20 ml) was added and the organic phase washedwith brine (2×50 ml), dried over sodium sulphate and evaporated. Theresidue was purified by flash chromatography (Biotage system) on silicagel using a 10 g SNAP column and cyclohexane to cyclohexane/ethylacetate 7:3 as eluents affording the title compound (0.05 g) as acolourless oil.

LC/MS: QC_(—)3_MIN: Rt=1.683 min.

Intermediate 1806[(3,3-diethyl-1H-isobenzofuran-4-yl)oxy]pyridin-3-amine

To a solution of 3,3-diethyl-1H-isobenzofuran-4-ol (Intermediate 179,0.03 g, 0.15 mmol) in dry DMF (3 ml) potassium carbonate (0.08 g, 0.6mmol) and then 2-Chloro-5-Nitropyridine (0.026 g, 0.17 mmoli) were addedand the reaction mixture was stirred for 2 hours at 80° C. After coolingthe reaction mixture was quenched with water (1 ml), diluted with brine(5 ml) and extracted with ethyl acetate (2×10 ml). The organic layer wasdried over sodium sulfate, filtered and evaporated. The residue wasdissolved in tetrahydrofuran (5 ml)/water (2.5 ml), iron (0.04 g, 0.75mmol) and then ammonium chloride (0.4 g, 0.75 mmol) were added and thereaction mixture was stirred overnight at room temperature. The catalystwas filtered off and the residue was diluted with an aqueous saturatedsolution of NaHCO₃ (5 ml) and extracted with ethyl acetate (3×10 ml).The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 10 g SNAP column and cyclohexane/ethylacetate 8:2 to cyclohexane/ethyl acetate 1:1 as eluents affording thetitle compound (0.025 g) as a light yellow solid.

LC/MS: QC_(—)3_MIN: Rt 1.735 min, 285 [M+H]+.

Intermediate 181[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy][tris(1-methylethyl)]silane

3,3-Dimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 50, 3.6 g,21.91 mmol) was dissolved in anhydrous THF (20.0 mL) and the colorlesssolution was cooled to 0° C. stirring under nitrogen. A 2M n-BuLisolution in cyclohexane (13.2 mL, 26.4 mmol) was added drop wise and theresulting yellow solution was stirred at 0° C. for 10 min.Triisopropylsislyltriflate (7.7 mL, 28.5 mmol) was added drop wise: thesolution discolored almost completely. This was allowed to warm to roomtemperature and stirred over night. Water (1.0 mL) was added to andvolatiles evaporated under reduced pressure. The residue was dissolvedin ethyl acetate and washed with brine three times. The organic layerwas dried over anhydrous Na₂SO₄ and evaporated to dryness to give yellowoil which was re-dissolved in TBME and washed twice with water. Theorganic solution was dried over Na₂SO₄ and evaporated to dryness to givethe title compound (7.4 g) as a yellow oil.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.94 (1H, t), 6.31-6.36 (1H, m), 6.29(1H, d), 4.14 (2H, s), 1.28-1.40 (9H, m), 1.09 (18H, d).

Intermediate 182[(7-bromo-3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy][tris(1-methylethyl])silane

[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy][tris(1-methylethyl)]silane(Intermediate 181, 7.4 g, 23.19 mmol) was dissolved in THF (70.0 mL).N-Bromosuccinimide (4.2 g, 23.88 mmol) was added dissolving in fewminutes. This mixture was stirred at room temperature for 3 hrs. MoreNBS (0.64 g, 3.48 mmol) was added and the reaction mixture was stirredat room temperature for a further hour. CCl₄ (50 mL) was added to thereaction mixture and the solution was evaporated to dryness. The residuewas re-suspended in CCl₄ and stirred at room temperature for 15 min. Thewhite solid was removed by filtration and the wet cake was washed withmore CCl₄. The CCl₄ was swapped with ethyl acetate and the organicsolution was washed three times with 2.5% w/w aqueous NaHCO₃ and finallywith water. The organic solution was dried on anhydrous Na₂SO₄ andevaporated to dryness to give the title compound (8.6 g) as a brown oil.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.14 (1H, d), 6.29 (1H, d), 4.24 (2H,s), 1.27-1.41 (9H, m), 1.08 (18H, d).

Intermediate 183tris(1-methylethyl)[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]silane

[(7-bromo-3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy][tris(1-methylethyl)]silane(Intermediate 182, 7.1 g, 17.72 mmol) was dissolved in anhydrous THF (72mL) and cooled to 0° C. Tetramethylethylenediamine (8.0 mL, 53.16 mmol)was added and the yellow solution was stirred at 0° C. for 10 min. Asolution of 1.6 M butyllithium in hexane (22.5 mL, 35.4 mmol) was addeddrop wise over 10 minutes and then stirred at 0° C. for 15 min. Methyliodide (11 mL, 177.2 mmol) was added drop wise over 6 min. The whitesolid was removed by filtration and the wet cake was washed in with THF.The combined organic layers were evaporated to dryness. The residue wasdissolved in ethyl acetate and washed twice with aqueous NaHCO₃ and oncewith water. The organic solution was dried on anhydrous Na₂SO₄ andevaporated to dryness. to give brown oil. The residue was purified byflash chromatography on silica gel using cyclohexane tocyclohexane/ethyl acetate 1:1 as eluents affording the title compound(3.6 g) as a brown oil.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.76 (1H, d), 6.20 (1H, d), 4.14 (2H,s), 2.02 (3H, s), 1.28-1.39 (9H, m), 1.09 (18H, d).

Intermediate 184 3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-ol

Tris(1-methylethyl)[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]silane(Intermediate 183, 3.6 g, 10.84 mmol) was dissolved in THF (36 mL) toobtain a dark yellow solution. TBAF (8.5 g, 32.5 mmol) was added and thereaction mixture was stirred overnight at room temperature. The solventwas removed under reduced pressure. The residue was dissolved in ethylacetate and washed with aqueous HCl, then aqueous NaHCO₃ and finallybrine. The organic solution was dried over Na₂SO₄ and evaporated todryness and the residue was purified by flash chromatography on silicagel using cyclohexane to cyclohexane/ethyl acetate 95:5 as eluentsaffording the title compound (1.69 g) as colorless oil.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.06 (1H, s), 6.65-6.69 (1H, m), 6.19(1H, d), 4.11 (2H, s), 1.99 (3H, s), 1.33 (6H, s).

Intermediate 1855-nitro-2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]pyridine

3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 184, 0.9 g,5.0 mmol) was dissolved in CH₃CN (5 mL) in the presence of2-chloro-5-nitropyridine (790 mg, 5.0 mmol) and K₂CO₃ (1.72 g, 12.5mmol) and the resulting suspension was heated to 60° C. for 1.5 hrs. Themixture was then cooled to room temperature and diluted with water andethyl acetate. Two phases were separated and the organic layer waswashed with brine, then dried over Na₂SO₄ and evaporated to dryness, Theresidue was purified by flash chromatography on silica gel usingcyclohexane to cyclohexane/ethyl acetate 90:10 as eluents affording thetitle compound (0.92 g) as yellowish solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.04 (1H, d), 8.61 (1H, dd), 7.24 (1H,d), 7.02 (1H, d), 6.54 (1H, d), 4.21 (2H, s), 2.14 (3H, s), 1.21 (6H,s). ¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 166.6, 158.7, 147.2, 144.8, 140.4,135.8, 130.2, 126.1, 116.7, 114.5, 111.0, 83.6, 42.2, 26.0, 14.4.

Intermediate 1866-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine

5-Nitro-2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]pyridine(Intermediate 185, 920 mg, 3.0 mmol) was dissolved in EtOH (13.5 mL) andstirred under hydrogen atmosphere (2 bar) in the presence of Pd/C 10%w/w (46 mg, 5% w/w) at room temperature for 30 minutes. The catalyst wasfiltered off, washed with THF and the resulting solution evaporated todryness to afford an orange solid. The crude product was crystallizedfrom MeOH to the title compound (565 mg) as a beige solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.51 (1H, d), 7.05 (1H, dd), 6.85 (1H,d), 6.69 (1H, d), 6.21 (1H, d), 5.04 (2H, br.s), 4.19 (2H, s), 2.08 (3H,s), 1.30 (6H, s). ¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 158.3, 154.2, 150.7,141.5, 132.2, 129.6, 125.3, 124.7, 113.9, 112.2, 111.8, 83.7, 42.2,26.0, 14.4.

Intermediate 187 1,1-dimethylethyl{(1R)-1-[({6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

6-{[3,3,7-Trimethyl-6-(trifluoromethoxy)-2,3-dihydro-1-benzofuran-4-yl]oxy}pyridin-3-amine(Intermediate 186, 405 mg, 1.27 mmol) was suspended in ethyl acetate (4mL). Triethylamine (0.44 ml, 3.175 mmol) was added followed by theaddition of (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoicacid (258 mg, 1.27 mmol). The resulting suspension was cooled to 0° C.and T3P 50% w/w solution in ethyl acetate (1.4 mmol) was added dropwise. The reaction mixture was stirred at 0° C. for 1 hour and thenwarmed to room temperature and stirred for a further hour. An aqueoussaturated solution of Na₂CO₃ was added and the mixture stirred for 10min. Two phases were separated and the organic layer was washed withwater and brine, dried over Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography on silica gel usingcyclohexane/ethyl acetate 80:20 to cyclohexane/ethyl acetate 70:30 aseluents affording the title compound (0.50 g) as white foam.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.08 and 10.03 (1H, br.s), 8.30 (1H,d), 8.03 (1H, dd), 7.00 (1H, d), 6.95-6.90 (2H, m), 6.36 (1H, d), 4.17(2H, s), 3.98-3.92 (1H, m), 2.10 (3H, s), 1.73-1.52 (2H, m), 1.36 and1.29 (9H, br.s), 1.23 (6H, s), 0.88 (3H, t). ¹³C-NMR (200 MHz, DMSO-d₆):δ ppm 171.4, 159.0, 158.5, 155.5, 148.9, 138.1, 131.4, 129.8, 125.8,115.1, 113.9, 110.7, 83.6, 78.0, 56.3, 42.2, 28.9, 26.0, 25.0, 20.7,14.4, 14.1, 10.5.

Intermediate 188(2R)-2-amino-N-{6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide

The 1,1-dimethylethyl{(1R)-1-[({6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Intermediate 187, 480 mg, 1.05 mmol) was dissolved in iso-propylacetate (5 mL) and HCl 5-6N in isopropanol (1 ml, 5.25 mmol) was added.The solution was stirred at room temperature for 1 hour and then heatedto ˜50-55° C. until complete conversion. The mixture was cooled to roomtemperature and treated with an aqueous saturated solution of NaHCO₃.Two phases were separated and the organic layer was washed with brine,dried over Na₂SO₄ and evaporated to dryness. The residue was purified byflash chromatography on silica gel using dichloromethane/methanol 95:5as eluents affording the title compound (0.31 g) as yellowish foam.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.36 (1H, d), 8.11 (1H, dd), 6.96-6.92(2H, m), 6.38 (1H, d), 4.19 (2H, s), 3.23 (1H, dd), 2.11 (3H, s),1.72-1.61 (1H, m), 1.53-1.43 (1H, m), 1.25 (6H, s), 0.90 (3H, t).¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 174.5, 159.0, 158.5, 148.9, 138.2,131.5, 131.4, 129.8, 125.7, 115.1, 113.9, 110.6, 83.6, 56.7, 42.2, 28.0,26.0, 14.4, 10.2.

Intermediate 1895-nitro-2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]pyrimidine

3,3,7-Trimethyl-2,3-dihydro-1-benzofuran-4-ol (Intermediate 184, 178 mg,1.0 mmol) and 2-chloro-5-nitropyrimidine (191.5 mg, 1.2 mmol) weredissolved in CH₃CN (3.0 mL) and K₂CO₃ (345.5 mg, 2.5 mmol) was added.The resulting suspension was heated to 40° C. and stirred for 1 hour.The reaction mixture was then diluted with water (50 mL) and ethylacetate (50 mL), The organic phase was collected, washed with brine (50mL) and dried over Na₂SO₄. The residue was purified by flashchromatography on silica gel using cyclohexane/ethyl acetate 97:3 aseluents affording the title compound (243 mg).

Intermediate 1902-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine

5-nitro-2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]pyrimidine(Intermediate 189, 243 mg, 0.81 mmol) was dissolved in THF (4 mL) andPalladium on charcoal (5 mol %, 85 mg) was added. The reaction mixturewas stirred under hydrogen atmosphere (3 bar) for 1 hour at roomtemperature. The catalyst was filtered on a pad of celite, washed withTHF and the resulting solution was concentrated under vacuum. Theresidue was diluted with ethyl acetate and water, the organic phasecollected, dried over Na₂SO₄ and evaporated to afford the title compound(220 mg) as colorless oil. The crude product, was used in the next stepwithout further purification.

MS_(—)2 (ESI): 272 [M+H]+

Intermediate 191 1,1-dimethylethyl{(1R)-1-[({2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate

2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinamine(Intermediate 190, 220 mg, 0.81 mmol) was dissolved in ethyl acetate (10mL) and of (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid(181.1 mg, 0.89 mmol) was added followed by the addition of Et₃N (0.35mL, 2.02 mmol). The resulting solution was cooled down to 5° C. and asolution of T3P 50% w/w in ethyl acetate (0.53 mL, 0.89 mmol) was addeddrop wise in 15 min. The reaction mixture was stirred for 30 min at 5°C. The reaction was quenched with water (50 mL) and ethyl acetate (50mL), two phases were separated and the organic layer was dried overNa₂SO₄ and concentrated under vacuum. The residue was purified by flashchromatography on silica gel using cyclohexane/ethyl acetate 60:40 aseluent affording the title compound (213 mg).

MS_(—)2 (61):457 [M+H]+.

Intermediate 192 7-(methoxymethoxy)-3H-isobenzofuran-1-one

4-Hydroxy-1,3-dihydro-2-benzofuran-1,3-dione (685 mg, 4 mmol) wasdissolved in dry THF (30 mL) at −78° C. K-Selectride 1M solution in THF(13 mL, 13 mmol) was added drop wise in 20 min then the mixture waswarmed from −78° C. to −30° C. over 3 hrs. The final mixture was pouredinto ethyl acetate (100 mL), brine (25 mL) and a 3M hydrochloric acidsolution (25 mL). The organic layer was collected, washed with brine andevaporated to dryness. The resulting compound was dissolved in MeOH (20mL) and treated under stirring with a 3M hydrochloric acid solution (10mL). The mixture was then diluted with ethyl acetate (100 mL) and brine(25 mL), the organic layer was collected and evaporated under vacuum.The residue was purified by flash chromatography on silica gel usingdichloromethane as eluent affording the phenol intermediate (430 mg).

The phenol intermediate (430 mg, 2.9 mmol) was dissolved in drydichloromethane (20 mL) at 0° C., DIPEA (5 mL, 5.5 mmol) was addedfollowed by a drop wise addition of chloromethylmethylether (0.44 mL,5.7 mmol) over 10 min. The resulting solution was stirred at 0° C. for30 minutes then at room temperature for 15 minutes. Dichloromethane waspartially evaporated and the obtained suspension was dissolved in ethylacetate (20 mL), washed with a 50/50 mixture of water and brine (2×10mL), dried over Na₂SO₄ and evaporated to give the title compound (549mg) as a pale yellow solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.68 (1H, m), 7.20 (2H, m), 5.36 (2H,s), 5.31 (2H, s), 3.41 (3H, s).

Intermediate 193 3,3-dimethyl-1H-isobenzofuran-4-ol

7-(methoxymethoxy)-3H-isobenzofuran-1-one (Intermediate 192, 550 mg, 2.8mmol) was dissolved at −70° C. in dry THF (150 mL). A 3M solution ofmethylmagnesium bromide in diethyl ether (5.6 mL, 16.8 mmol) was addeddrop wise in 30 minutes and the obtained mixture stirred for 30 minutesat −70° C. and then 30 minutes at room temperature. The reaction mixturewas poured into ethyl acetate (100 mL) and aqueous saturated solution ofammonium chloride (50 mL) at 0° C. The organic layer was collected,washed with an aqueous saturated solution of ammonium chloride (50 mL),brine (50 mL) and evaporated to give a yellow oil that was dissolved inacetonitrile (15 mL) and treated with sulphuric acid (0.15 mL).Acetonitrile was replaced with methyl alcohol (15 mL) and the resultingsolution treated with p-toluenesulfonic acid (100 mg). The solution washeated to 60° C. and stirred for 2 hours. The solvent was evaporated andthe residue dissolved in ethyl acetate (30 mL), washed twice with anaqueous saturated solution of sodium bicarbonate (10 mL) and then withan aqueous diluted solution of hydrochloric acid, dried over Na₂SO₄, andevaporated to dryness. The residue was purified by flash chromatographyon silica gel using cyclohexane/ethyl acetate 90:10 as eluent affordingthe title compound (210 mg) as white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ ppm 9.51 (1H, s), 7.00-7.10 (1H, m),6.60-6.70 (2H, m), 4.89 (2H, s), 1.45 (6H, s).

Intermediate 194(5R)-3-(6-chloro-3-pyridinyl)-5-ethyl-5-methyl-2,4-imidazolidinedione

6-Chloropyridin-3-amine (3.0 g, 23.3 mmol) was dissolved in a 3:1 v/vmixture of CH₃CN/ethyl acetate (20 mL) and (2R)-2-amino-2-methylbutanoicacid hydrochloride (3.97 g, 25.63 mmol) was added, followed by a 50% w/wsolution in ethyl acetate of T3P (15.3 mL, 25.63). The mixture washeated to 60° C. for 2 hrs, then quenched with NaOH 3N while the pH was˜10 and then diluted with ethyl acetate (100 mL). The organic phase wascollected, dried over Na₂SO₄ and concentrated under vacuum to a finalvolume of ˜15 mL. The solution was cooled down to 0-5° C. and Et₃N (11.4ml, 81.9 mmol) was added. A solution of triphosgene (2.76 g, 6.96 mmol)in 10 mL of ethyl acetate was added drop wise in 15 min, keeping theinternal temperature below 5° C. The mixture was stirred at 5° C. for 30minutes then quenched with water (100 mL) and finally diluted withadditional ethyl acetate (100 mL). Two phases were separated and theorganic layer was washed with water and brine, dried over Na₂SO₄ andconcentrated under vacuum. The crude product was suspended in 15 mL ofethyl acetate followed by the drop wise addition of 65 mL of n-heptane.The resulting suspension was stirred at room temperature for 2 hrs,filtered, and the cake washed with a 2:8 v/v mixture of ethylacetate/n-heptane (2×10 mL) before being dried at 40° C. for 18 hrsaffording the title compound (3.5 g) as white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.68 (1H, s), 8.47 (1H, d), 7.95-7.89(m, 1H), 7.68-7.63 (1H, m), 1.83-1.58 (2H, m), 1.38 (3H, s), 0.85 (3H,t).

Intermediate 195 (2,2-difluoro-1,3-benzodioxol-4-yl)boronic acid

2,2-Difluoro-1,3-benzodioxole (960 mg, 6.1 mmol) was dissolved in THF (8mL) and cyclohexane (4 mL) and the resulting solution cooled to −78° C.sec-BuLi 1.4M solution in cyclohexane (4.3 mL, 6.1 mmol) was addeddropwise and the reaction mixture stirred for 1.5 hours at −78° C.Trimethylborate (694 mg, 6.75 mmol) was added and the mixture wasallowed to warm slowly to −30° C. The reaction mixture was quenched witha 2N solution of HCl and diluted with ethyl acetate. Two phases wereseparated and the organic layer was washed twice with brine, dried overNa₂SO₄ and evaporated to dryness affording the title compound as yellowoil which was used in the next step without further purification.

¹H NMR (400 MHz, DMSO-d₆+D₂O): δ ppm 7.39 (1H, dd), 7.34 (1H, dd), 7.14(t, 1H, J=7.90 Hz). ¹⁹F-NMR (376 MHz, DMSO-d₆+D₂O): δ ppm −48.92.¹³C-NMR (200 MHz, DMSO-d₆+D₂O): δ ppm 147.3, 142.8, 131.6 (t, J=250.7Hz), 130.1, 124.3, 112.0

Intermediate 196

(2,2-difluoro-7-methyl-1,3-benzodioxol-4-yl)boronic acid

(2,2-difluoro-1,3-benzodioxol-4-yl)boronic acid (Intermediate 195, crudematerial) was dissolved in THF (20 mL) and the resulting solution cooleddown to −78° C. sec-BuLi 1.4M solution in cyclohexane (17.4 ml, 24.36mmol) was added dropwise and the reaction mixture was stirred for 1.5hours at −78° C. Methyl iodide (4.6 ml, 73 mmol) was then added and thereaction mixture was stirred for 2 hours while the temperature wasallowed to reach room temperature. The reaction was quenched by additionof an aqueous 2N solution of HCl and diluted with ethyl acetate. Theorganic layer was collected and then washed twice with brine, dried overNa₂SO₄ and evaporated to dryness. Crystallization from n-heptaneafforded the title compound (150 mg) as white solid.

¹H NMR (400 MHz, DMSO-d₆+D₂O): δ ppm 7.30 (1H, d), 6.68 (1H, d), 2.25(s, 3H). ¹⁹F-NMR (376 MHz, DMSO-d₆+D₂O): δ ppm −48.55. ¹³C-NMR (200 MHz,DMSO-d₆+D₂O): δ ppm 152.5, 147.1, 141.5, 131.6 (t, J=250.0 Hz), 129.9,125.8, 122.7, 110.1, 14.6.

Intermediate 197 2,2-difluoro-7-methyl-1,3-benzodioxol-4-ol

(2,2-difluoro-7-methyl-1,3-benzodioxol-4-yl)boronic acid (Intermediate196, 150 mg, 1.28 mmol) was dissolved in THF (1.5 mL) and a 30% w/waqueous solution of H₂O₂ (2.56 mmol) and NaOH (51 mg, 1.28 mmol) wereadded and the reaction mixture stirred for 2 days at room temperature.The reaction was quenched with a 2N aqueous solution of HCl and dilutedwith ethyl acetate. Two phases were separated and the organic layer waswashed twice with brine, dried over Na₂SO₄ and evaporated to dryness,affording the title compound (140 mg) as yellow oil.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.31 (1H, s), 6.83 (1H, d), 6.63 (1H,d), 2.17 (3H, s). ¹⁹F-NMR (376 MHz, DMSO-d₆): δ ppm −48.68. ¹³C-NMR (200MHz, DMSO-d₆): δ ppm 142.3, 139.1, 131.4 (t, J=251.9 Hz), 129.9, 125.6,112.8, 110.0, 13.2.

Intermediate 198 2,2-difluoro-1,3-benzodioxol-4-ol

2,2-Difluoro-1,3-benzodioxole (320 mg, 2.05 mmol) was dissolved in THF(2.5 mL) and cyclohexane (1.2 mL) and the resulting solution cooled downto −78° C. sec-BuLi 2M solution in cyclohexane (1.025 ml, 2.05 mmol) wasadded drop wise and the reaction mixture stirred for 2 hours at −78° C.Trimethylborate (230 mg, 2.25 mmol) was added and the mixture wasallowed to warm slowly to room temperature. A 30% w/w aqueous solutionof H₂O₂ (4.1 mmol) and NaOH (82 mg, 2.05 mmol) were added and thereaction mixture stirred for 18 hours at room temperature. The reactionwas quenched with a 2N aqueous solution of HCl and diluted with ethylacetate. Two phases were separated and the organic layer was washedtwice with brine, dried over Na₂SO₄ and evaporated to dryness, affordingthe title compound (340 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 6.93 (1H, t), 6.69 (1H, d), 6.65 (1H, d).¹⁹F-NMR (376 MHz, CDCl₃): 6 ppm −49.86. ¹³C-NMR (200 MHz, CDCl₃): δ ppm144.8, 139.6, 131.5 (t, J=255.1 Hz), 131.2, 123.9, 112.7, 101.8

Intermediate 199 2-[2-amino-6-(methyloxy)phenyl]-2-propanol

1-(2-Amino-6-methoxyphenyl)ethanone (500 mg, 3.03 mmol) was dissolved inTHF (7.5 mL) and cooled to 0° C. A 3M solution of Methyl magnesiumbromide in Et₂O (2.12 ml, 6.36 mmol) was added drop wise keeping thetemperature below 10° C. The reaction was quenched with an aqueoussaturated solution of NH₄Cl (7.5 mL) keeping the temperature below 15°C. The mixture was diluted with water and ethyl acetate, two phasesseparated and the organic layer was washed twice with brine, dried overNa₂SO₄ and evaporated to dryness affording the title compound (500 mg)as light orange solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 6.79 (1H, t), 6.19 (1H, dd), 6.14 (1H,dd), 5.76 (2H, br.s), 5.26 (1H, br.s), 3.64 (3H, s), 1.55 (6H, s).¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 157.4, 148.5, 126.8, 118.1, 110.5,100.2, 74.2, 55.3, 30.7.

Intermediate 200N-[2-(1-hydroxy-1-methylethyl)-3-(methyloxy)phenyl]acetamide

2-(2-Amino-6-methoxyphenyl)propan-2-ol (Intermediate 199, 500 mg, 2.76mmol) was dissolved in DCM (10 mL) and triethylamine (0.770 ml, 5.52mmol) was added. The solution was cooled to 0° C. and treated withacetyl chloride (0.2 ml, 2.76 mmol) in a drop wise fashion. At the endof the addition, complete conversion was reached. The mixture wastreated with an aqueous saturated solution of NH₄Cl, two phases wereseparated and the organic layer was washed with an aqueous saturatedsolution of NaHCO₃ and brine, then dried over Na₂SO₄ and evaporated todryness. The crude was purified by re-slurry in tert-butyl methyl etherisolating 490 mg of the title compound as white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 11.53 (1H, br.s), 7.91 (1H, d), 7.11(1H, t), 6.71 (1H, d), 6.25 (1H, br.s), 3.74 (3H, s), 2.50 (1H, br.s),2.00 (3H, s), 1.61 (6H, s). ¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 167.0,156.6, 138.5, 127.0, 123.1, 114.3, 107.6, 75.2, 55.7, 30.8, 25.2.

Intermediate 201 2,4,4-trimethyl-5-(methyloxy)-4H-3,1-benzoxazine

N-[2-(2-hydroxypropan-2-yl)-3-methoxyphenyl]acetamide (Intermediate 200,470 mg, 2.10 mmol) was added to hot polyphosphoric acid and heated at110° C. for 1 hour. The mixture was cooled to room temperature andquenched with water. Solid Na₂CO₃ was added while the pH was allowed toreach ˜8-9. Water and DCM were added and two phases were separated. Thecombined organic layers were washed twice with brine, dried over Na₂SO₄and evaporated to dryness, to obtain the title compound (342 mg) as anoil which was used in the next step without further purifications.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.17 (1H, t), 6.83 (1H, dd), 6.63 (1H,dd), 3.78 (3H, s), 1.98 (3H, s), 1.60 (6H, s). ¹³C-NMR (200 MHz,DMSO-d₆): δ ppm 158.4, 154.7, 139.0, 128.6, 117.0 (2C), 109.7, 77.8,55.7, 28.5, 21.3.

Intermediate 202 2,4,4-trimethyl-4H-3,1-benzoxazin-5-ol

5-Methoxy-2,4,4-trimethyl-4H-3,1-benzoxazine (Intermediate 201, 342 mg,1.67 mmol) was dissolved in DCM (7 mL) and 1M BBr₃ solution in DCM (1.67ml, 1.67 mmol) was added. After 1.5 h at room temperature the mixturewas heated to reflux and after 6 hours at reflux, some more 1M BBr₃solution in DCM (1.67 ml, 1.67 mmol) was added leaving the mixture atreflux overnight. The reaction mixture was quenched with an aqueoussaturated solution of NaHCO₃ while the pH was allowed to become basic.Two phases were separated and the organic layer was washed with brine,dried over Na₂SO₄ and evaporated to dryness, to obtain the titlecompound (330 mg) as foam.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.62 (1, s), 6.98 (1H, t), 6.62 (1H,dd), 6.46 (1H, dd), 1.96 (3H, s), 1.62 (6H, s). ¹³C-NMR (200 MHz,DMSO-d₆): δ ppm 157.9, 152.6, 139.2, 128.2, 115.4, 115.3, 113.9, 77.8,28.3, 21.3.

Intermediate 2032,4,4-trimethyl-5-[(5-nitro-2-pyridyl)oxy]-3,1-benzoxazine

To a suspension of 2,4,4-trimethyl-4H-3,1-benzoxazin-5-ol (Intermediate202, 500 mg, 2.61 mmol) and 2-chloro-5-nitropyridine (410 mg, 2.58 mmol)in dry DMF (4 mL) potassium carbonate (400 mg, 2.89 mmol) was added andthe resulting mixture was heated in a MW apparatus at 70° C. for 40 min.The mixture was diluted with water and ethyl acetate, phases wereseparated and the aqueous was back-extracted with ethyl acetate (2×20mL). The combined organic layers were washed with brine (2×20 mL), driedover Na₂SO₄, filtered and concentrated and the residue was purified byflash chromatography on silica gel using cyclohexane/ethyl acetate from70:30 to 50:50 as eluents affording the title compound (225 mg) asyellow foam.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.05 (1H, d), 8.63 (1H, dd), 7.21-7.40(2H, m), 6.84-7.02 (2H, m), 2.01 (3H, s), 1.50 (6H, s).

Intermediate 2046-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]pyridin-3-amine

To a solution of2,4,4-trimethyl-5-[(5-nitro-2-pyridyl)oxy]-3,1-benzoxazine (Intermediate203, 220 mg, 0.70 mmol) in EtOH (3 mL) palladium on carbon 10% w/w (25mg) was added and the resulting mixture was stirred for 40 minutes underhydrogen atmosphere (2 bar) at room temperature. The catalyst wasfiltered off and washed with EtOH (3×10 mL). The filtrate wasconcentrated to the title compound (185 mg) as greenish solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.51 (1H, d), 7.12 (1H, t), 7.05 (1H,dd), 6.75 (2H, d), 6.55 (1H, d), 5.07 (2H, s), 1.99 (3H, s), 1.58 (6H,s).

Intermediate 205(2R)-2-amino-2-methyl-N-[6-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]-3-pyridyl]butanamide

To a suspension of6-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]pyridin-3-amine(Intermediate 204, 185 mg, 0.65 mmol) and (R)-2-amino-2-methyl-butanoicacid hydrochloride (100 mg, 0.67 mmol) in ethyl acetate/MeCN (2 mL, 1:3v/v mixture), a 50% w/w solution in ethyl acetate of T3P (0.43 mL) wasadded drop wise at 0° C. The mixture was then heated at 60° C. for 3hours and at 80° C. for 4.5 hours. The mixture was cooled to roomtemperature diluted with water (5 mL) and ethyl acetate (10 mL), Twophases were separated and the aqueous one was treated with a saturatedsolution of NaHCO₃ (pH=8) and back-extracted with ethyl acetate (2×10mL). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated and the residue was purified by flash chromatography onsilica gel using cyclohexane/ethyl acetate 20:80 as eluent affording thetitle compound (143 mg) as a yellow foam.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.39 (1H, d), 8.17 (1H, dd), 7.20 (1H,t), 7.01 (1H, d), 6.84 (1H, d), 6.73 (1H, d), 2.00 (3H, s), 1.69 (1H,m), 1.43-1.57 (7H, m), 1.21 (3H, s), 0.80 (3H, t).

Intermediate 2062-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-nitropyridine

In a microwave vial, 2-chloro-5-nitropyridine (97 mg, 0.609 mmol) wasdissolved in 3 mL of dimethylformamide.2,2-dimethyl-2,3-dihydro-1-benzofuran-4-ol (100 mg, 0.609 mmol) andpotassium carbonate (253 mg, 1.827 mmol) were added. The reactionmixture was heated under microwave irradiation for 1 hour at 110 C. Thereaction mixture was filtered. The filtrated solid was washed withdichloromethane (5 ml). The volatiles were evaporated under vacuum. Thecrude compound was dissolved in dichloromethane (8 ml) and brine wasadded (8 ml). The compound was extracted 2 times with dichloromethane(2×8 ml) and 2 times with ethylacetate (2×8 ml). Combined organic layerswere dried over sodium sulphate and evaporated. The residue was purifiedby silica gel chromatography (Companion system, 12 g Si cartridge) withcyclohexane/ethyl acetate from 100:0 to 80:20 as eluents affording thetitle compound (120 mg).

UPLC_ipqc: 1.20 min, 287 [M+H]+.

Intermediate 2076-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine

Fe powder (112 mg, 2.009 mmol) was added to a solution of2-[(2,2-dimethyl-2,3-dihydro-h-benzofuran-4-yl)oxy]-5-nitropyridine(Intermediate 206, 115 mg 0.402 mmol) in a mixture THF/water (9 ml/3 ml)followed by the addition of ammonium chloride (107 mg 2.009 mmol). Thereaction mixture was stirred overnight at room temperature. The catalystwas filtered off and the solution was diluted with an aqueous saturatedaqueous solution of NaHCO₃ (10 ml) and ethyl acetate (15 ml). Two phaseswere separated and the aqueous layer was extracted twice with ethylacetate (2×15 ml). Combined organic layers were dried over Na₂SO₄,filtered and evaporated. The residue was purified by flashchromatography (Companion system, 12 g silica gel cartridge) usingcyclohexane/ethyl acetate from 80:20 to 50:50 as eluents affording thetitle compound (95 mg).

UPLC_ipqc: 0.87 min, 257 [M+H]+.

Intermediate 208 tert-butylN-[(1R)-1-[[6-[(2,2-dimethyl-3H-benzofuran-4-yl)oxy]-3-pyridyl]carbamoyl]propyl]carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (17.84 mg,0.088 mmol) in dry N,Ndimethylformamide (DMF) (1 mL), DIPEA (25.6 μl,0.146 mmol) and then HATU (37.8 mg, 0.099 mmol) were added and thereaction mixture was stirred for 15 minutes at room temperature underargon. Then6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinamine(Intermediate 207, 15 mg, 0.059 mmol) was added and the reaction mixturewas left overnight under stirring at 35° C. under argon. The reactionmixture was evaporated. Brine (4 ml) was added and it was extracted 3times with ethyl acetate (3×5 ml). Combined organic layers were driedover Na₂SO₄, filtered and evaporated and the residue was purified byflash chromatography (Companion system, 12 g silica cartridge) withcyclohexane/ethyl acetate as eluents from 100:0 to 70:30 affording thetitle compound (18 mg).

UPLC_ipqc: 1.21 min, 442 [M+H]+.

Intermediate 209((2R)-2-amino-N-{6-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide)

To a solution of tert-butylN-[(1R)-1-[[6-[(2,2-dimethyl-3H-benzofuran-4-yl)oxy]-3-pyridyl]carbamoyl]propyl]carbamate(Intermediate 208, 14 mg, 0.032 mmol) in dry dichloromethane (1 ml) at0° C., TFA (98 μl, 1.268 mmol) was slowly added and the reaction mixturewas stirred for 4 hours at the same temperature. Some moredichloromethane (4 ml) was added to the reaction mixture. An aqueoussaturated aqueous solution of NaHCO₃ was then added while the pH wasallowed to reach ˜8. Two phases were separated and the aqueous one wasfurther extracted with DCM (3×3 ml). Combined organic layers were driedover Na₂SO₄, filtrated and evaporated affording the title compound (10mg).

UPLC_ipqc: 0.73 min, 342 [M+H]+.

Intermediate 210 5-(methoxymethoxy)-4-methyl-chromane

In a 50 mL round-bottomed flask2-(3-hydroxy-1-methyl-propyl)-3-(methoxymethoxy)phenol (56.3 mg, 0.249mmol) was dissolved in Tetrahydrofuran (THF) to give a colourlesssolution. Triphenylphosphine (59.4 mg, 0.226 mmol) was added and thereaction mixture was stirred until complete solubilization oftriphenylphosphine. DIAD (45.8 mg, 0.226 mmol) was added and thereaction mixture was stirred. Additional Triphenylphosphine (59.4 mg,0.226 mmol) and DIAD (45.8 mg, 0.226 mmol) were added. The reactionmixture evaporated in vacuo and the residue was purified by flashchromatography (Biotage SP1) on silica gel using a 10 g SNAP silicacartridge as column and cyclohexane/ethyl acetate 10:1 as eluentsaffording the title compound (50.3 mg) as colorless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.03 (1H, t), 6.63 (1H, d), 6.52 (1H, d),5.22 (2H, dd), 4.20-4.28 (1H, m), 4.09-4.19 (1H, m), 3.51 (3H, s),3.09-3.23 (1H, m), 1.96-2.17 (1H, m), 1.61-1.74 (1H, m), 1.31 (3H, d).

Intermediate 211 4-methylchroman-5-ol

In a 50 mL round-bottomed flask 5-(methoxymethoxy)-4-methyl-chromane(Intermediate 210, 50.3 mg, 0.229 mmol) was dissolved in Methanol (4 mL)to give a pale yellow solution. A 2M aqueous solution of HCl (0.100 mL,0.200 mmol) was added. The reaction mixture was stirred at 50° C.Sequential addition of 2M/H₂O solution of HCl (0.100 mL, 0.200 mmol)were added until completion of reaction. The reaction mixture wasquenched with 10 mL of water and diluted with 25 mL of DCM. Phases wereseparated through a phase separator cartridge. The organic layer wasevaporated in vacuo affording the title compound (38.7 mg) as orangeoil.

¹H NMR (400 MHz, CDCl₃) δ ppm 6.95 (1H, t), 6.44 (1H, d), 6.32 (1H, dd),4.87 (1H, br. s.), 4.20-4.31 (1H, m), 4.07-4.19 (1H, m), 3.05-3.15 (1H,m), 2.02-2.19 (1H, m), 1.66-1.74 (1H, m), 1.32 (3H, d).

Intermediate 212 2-(4-methylchroman-5-yl)oxy-5-nitro-pyridine

In a 0.5-2 ml Microwave vial 4-methylchroman-5-ol (Intermediate 211,38.7 mg, 0.212 mmol) K₂CO₃ (88 mg, 0.636 mmol) and2-chloro-5-nitropyridine (33.6 mg, 0.212 mmol) were dissolved inN,N-dimethylformamide (DMF) (2 mL) to give a light brown solution. Thereaction vessel was sealed and heated under microwave irradiation at110° C. for 1 hour. After cooling the reaction was quenched with 5 mL ofwater and diluted with 10 mL of ethyl acetate. Phases were separated bya separating funnel. The aqueous phase was extracted with 3×10 mL ofethyl acetate. The collected organic layer was dried using a hydrophobicfrit and evaporated in vacuo to give the title compound (22.9 mg) as acolorless oil.

UPLC_B: 0.94 min, 287 [M+H]+.

Intermediate 213 6-(4-methylchroman-5-yl)oxypyridin-3-amine

In a 50 mL round-bottomed flask2-(4-methylchroman-5-yl)oxy-5-nitro-pyridine (Intermediate 212, 22.9 mg,0.08 mmol) was dissolved in Ethanol (10 mL) to give a pale yellowsolution. Pd/C (17.88 mg, 0.017 mmol) and hydrazine hydrate (0.4 mL,4.15 mmol) were added. The reaction mixture was stirred at 90° C. Thereaction mixture was filtered and evaporated in vacuo to give the titlecompound (22.9 mg) as a pale yellow oil.

UPLC_B: 0.65 min, 257 [M+H]+.

Intermediate 214 tert-butylN-[1,1-dimethyl-2-[[6-(4-methylchroman-5-yl)oxy-3-pyridyl]amino]-2-oxo-ethyl]carbamate

In a 8 mL vial the 6-(4-methylchroman-5-yl)oxypyridin-3-amine(Intermediate 213, 22.9 mg, 0.089 mmol) was dissolved inN,N-Dimethylformamide (3 mL) to give a pale yellow solution. DIPEA(0.069 mL, 0.394 mmol),N-{[(1,1-dimethylethyl)oxy]carbonyl}-2-methylalanine (60.0 mg, 0.295mmol) and HATU (112 mg, 0.295 mmol) were added. The reaction mixture wasshaken at 60° C. for 1 hour. The reaction mixture was evaporated invacuo and the residue was purified by flash chromatography on silica gelusing a column SNAP 25 g and cyclohexane/ethyl acetate from 3:1 to 1:2as eluents affording the title compound (63.3 mg) as a colorless oil.

UPLC_B: 0.91 min, 442 [M+H]+.

Intermediate 2152-amino-2-methyl-N-[6-(4-methylchroman-5-yl)oxy-3-pyridyl]propanamide

In a 50 mL round-bottomed flask tert-butylN-[1,1-dimethyl-2-[[6-(4-methylchroman-5-yl)oxy-3-pyridyl]amino]-2-oxo-ethyl]carbamate(Intermediate 214, 63.3 mg, 0.093 mmol), was dissolved inDichloromethane (DCM) (3 mL) to give a pale yellow solution. Thereaction mixture was cooled at 0° C. and TFA (3 mL, 38.9 mmol) wasadded. The reaction mixture was stirred at 0° C. for 2 hours. Thereaction mixture was evaporated in vacuo to give the crude product as apale yellow oil. The residue was charged on a 2 g SCX cartridge. It wasthen flushed with 40 mL of MeOH followed by 40 mL of 2M solution ofammonia in MeOH. The Ammonia eluate was evaporated in vacuo to give thetitle compound (22.9 mg) as a colourless oil.

UPLC_B: 0.77 min, 342 [M+H]+.

Intermediate 216 2-(4-methylchroman-5-yl)oxy-5-nitro-pyrimidine

4-methyl-3,4-dihydro-2Hchromen-5-ol (Intermediate 211, 111 mg, 0.676mmol) was dissolved in 5.0 mL of DMF. K₂CO₃ (140 mg, 1.01 mmol) and2-chloro-5-nitropyrimidine (162 mg, 1.01 mmol) were added and thereaction mixture was stirred for 1 hour at room temperature. DMF wasthen evaporated under high vacuum and the residue was purified by flashchromatography on silica gel using cyclohexane/ethyl acetate from 1:0 to7:3 as eluents affording the title compound (192 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 9.36 (2H, s), 7.21 (1H, t), 6.85 (1H, d),6.68 (1H, d), 4.15-4.35 (2H, m), 2.90-3.02 (1H, m), 2.08-2.20 (1H, m),1.65-1.75 (1H, m), 1.29 (3H, d).

Intermediate 2172-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinamine

2-[(4-methyl-3,4-dihydro-2Hchromen-5-yl)oxy]-5-nitropyrimidine(Intermediate 216, 192 mg, 0.668 mmol) was dissolved in 9.0 mL of a 2/1THF/water solution. Iron (187 mg, 3.34 mmol) and ammonium chloride (179mg, 3.34 mmol) were added and the reaction mixture was stirred for 24 hat room temperature. After dilution with AcOEt and filtration over acelite pad (washing with AcOEt), the organic phase was washed (twotimes) with an aqueous saturated solution of NaHCO₃. The organic layerwas dried over Na₂SO₄, filtered and evaporated and the residue waspurified by flash chromatography on silica gel using cyclohexane/ethylacetate from 1:0 to 0:1 as eluents affording the title compound (144mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.11 (2H, s), 7.13 (1H, t), 6.73 (1H, d),6.60 (1H, d), 4.12-4.30 (2H, m), 3.52 (1H, br.s), 3.05-3.45 (1H, m),2.08-2.18 (1H, m), 1.63-1.73 (1H, m), 1.31 (3H, d).

Intermediate 218 1,1-dimethylethyl{(1R)-1-methyl-1-[({2-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate

2-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinamine(Intermediate 217, 144 mg, 0.56 mmol) was dissolved in toluene (8.0 mL)and S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 86 mg, 0.28 mmol) was added. The reaction mixture wasstirred at 140° C. for 20 minutes. Additional S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 134 mg, 0.43 mmol) was added and the mixture wasstirred at 140° C. for 15 minutes. Additional S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 100 mg, 0.32 mmol) was added and the reaction mixturewas stirred at 50° C. overnight and at 80° C. and for 4 hours.Additional S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 100 mg, 0.32 mmol) was added and the reaction mixturewas stirred at 80° C. for 20 hours. Additional S-2-pyridinyl(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-methylbutanethioate(Intermediate 139, 50 mg, 0.16 mmol) was added and the reaction mixturewas stirred for additional 30 hours at 80° C. After cooling volatileswere removed and the residue was purified by flash chromatography(Biotage SP1) on silica gel using a 10 g SNAP silica cartridge as columnand cyclohexane/ethyl acetate from 10:0 to 1:1 as eluents affording thetitle compound (100 mg).

UPLC_B: 1.13 min, 457 [M+H]+.

Intermediate 219N1-{2-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinyl}-D-isovalinamide

To a solution of 1,1-dimethylethyl{(1R)-1-methyl-1-[({2-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate(Intermediate 218, 100 mg, 0.219 mmol) in dry Dichloromethane (5 mL),cooled to 0° C., TFA (1 mL, 12.98 mmol) was added dropwise. The reactionmixture was stirred at that temperature for 3 hours, then it was allowedto reach the room temperature and stirred at that temperature for 2hours. Volatiles were evaporated and the residue was diluted with DCM(10 mL) and washed with an aqueous saturated solution of NaHCO3 (10 mL).The organic layer was separated, dried (Na₂SO₄), filtered and evaporatedto afford the title compound (78 mg) as white solid that was used in thenext step without further purification.

UPLC_B: 0.68 min, 357 [M+H]+.

Intermediate 2205-{[(methyloxy)methyl]oxy}-2,3-dihydrospiro[chromene-4,1′-cyclopropane]

To a solution of 2,4,6-trichlorophenol (307 mg, 1.557 mmol) in 12.0 mLof DCM, at −40° C., 1M solution in hexane of diethylzinc (1.557 mL,1.557 mmol) was added. After stirring at that temperature for 15minutes, CH₂I₂ (0.126 mL, 1.557 mmol) was added. After stirring foradditional 15 minutes,4-methylidene-5-{[(methyloxy)methyl]oxy}-3,4-dihydro-2H-chromene (169mg, 0.819 mmol) dissolved in 3.0 mL of DCM was added. The cold bath wasremoved and the mixture was stirred at room temperature for 2 hours. Themixture was diluted with DCM, washed with a 10% aqueous solution of HCl2 times, then with an aqueous saturated solution of NaHCO₃ 2 times, anaqueous saturated solution of Na₂SO₃ 2 times and brine (2 times). Theorganic layer was then dried over Na₂OS₄, filtered and evaporated andthe residue was purified by flash chromatography (Biotage SP1) on silicagel using cyclohexane/ethyl acetate from 100:0 to 95:5. Collectedresidue was dissolved in DCM and washed (two times) with KOH (30% aqsol). The organic layer was dried over sodium sulphate, filtered andevaporated to afford the title compound (145 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 6.96 (1H, t), 6.52-6.62 (2H, m), 5.05-5.15(2H, m), 4.22 (2H, dd), 3.47 (3H, s), 1.80-1.86 (2H, m), 1.72-1.80 (2H,m), 0.51-0.61 (2H, m).

Intermediate 221 2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-ol

5-{[(methyloxy)methyl]oxy}-2,3-dihydrospiro[chromene-4,1′-cyclopropane](Intermediate 220, 145 mg, 0.658 mmol) was dissolved in MeOH (6.0 mL)and a 2N aqueous solution of HCl (0.494 mL, 0.99 mmol) was added and thereaction mixture was stirred at 50° C. overnight. After addition ofwater, MeOH was removed under vacuum and the aqueous phase was extractedwith ethyl acetate (three times). Combined organic layers were driedover Na₂SO₄, filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using cyclohexane tocyclohexane/ethyl acetate 9:1 as eluents affording the title compound(64 mg).

UPLC_B: 0.93 min, 177 [M+H]+.

Intermediate 2222-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-nitropyrimidine

To a solution of 2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-ol(Intermediate 221, 63 mg, 0.358 mmol) in DMF (3 ml) K₂CO₃ (74.1 mg,0.536 mmol) and 2-chloro-5-nitropyrimidine (86.0 mg, 0.536 mmol) wereadded and the reaction mixture was stirred at rt for 1 h. DMF was thenevaporated under high vacuum, water was added and the reaction mixturewas extracted with AcOEt (three times). Collected organic layers weredried over Na₂SO₄, filtered and evaporated. The residue was purified byflash chromatography (Biotage system) on silica gel using cyclohexane tocyclohexane/ethyl acetate 9:1 as eluents affording the title compound(91 mg).

UPLC_B: 1.10 min, 300 [M+H]+.

Intermediate 2232-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinamine

2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-nitropyrimidine(Intermediate 222, 91 mg, 0.304 mmol) was dissolved in 9.0 mL of a 2/1THF/water solution. Then iron (85 mg, 1.52 mmol) and ammonium chloride(81 mg, 1.52 mmol) were added and the reaction mixture was stirred atroom temperature for 10 h. After dilution with AcOEt and filtration overa celite pad (washing with AcOEt), the organic phase was washed (twotimes) with an aqueous saturated solution of NaHCO₃. The organic layerwas dried over Na₂SO₄, filtered and evaporated. The residue was purifiedby flash chromatography (Biotage system) on silica gel using cyclohexaneto cyclohexane/ethyl acetate 1:1 as eluents affording the title compound(52 mg).

UPLC_B: 0.82 min, 270 [M+H]+.

Intermediate 2241,1-dimethylethyl[(1R)-1-({[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]amino}carbonyl)-1-methylpropyl]carbamate

To a solution of (2R)-2-(tert-butoxycarbonylamino)-2-methyl-butanoicacid (105 mg, 0.483 mmol) in dry N,N-Dimethylformamide (2 mL) DIPEA(0.101 mL, 0.579 mmol) and HATU (184 mg, 0.483 mmol) were added. Thereaction mixture was stirred at r.t. for 15 minutes, then it was addedto a solution of2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinamine(Intermediate 223, 52 mg, 0.193 mmol) in dry DMF (0.5 mL). The mixturewas heated at 40° C. overnight, it was then warmed to 60° C. and stirredat that temperature for 4 hours. After cooling the reaction was quenchedwith water (5 mL) and extracted with ethyl acetate (3×5 mL). Combinedorganic layers were washed with brine (3×5 mL), dried (Na₂SO₄), filteredand evaporated. The residue was purified by flash chromatography onsilica gel using a column SNAP 25 g and toluene to toluene/ethyl acetate60:40 as eluents affording the title compound (10 mg) as a white solid.

UPLC_B: 1.15 min, 469 [M+H]+.

Intermediate 225N1-[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]-D-isovalinamide

To a solution of1,1-dimethylethyl[(1R)-1-({[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]amino}carbonyl)-1-methylpropyl]carbamate(Intermediate 224, 5 mg, 10.67 μmol) in dry Dichloromethane (1 mL)cooled to 0° C. TFA (0.822 μL, 10.67 μmol) was added dropwise. Thereaction mixture was stirred for 2 hours at the same temperature. Thereaction was allowed to reach room temperature then the volatiles wereevaporated. The residue was diluted with DCM (2 mL), and washed withNaHCO₃ sat. sol. (7 mL). The organic layer was dried (Na₂SO₄), filteredand evaporated to afford the title compound (4 mg) as a yellow oil thatwas used in the next step without further purification.

UPLC_B: 0.71 min, 369 [M+H]+.

Intermediate 2266-(1,1a,2,7btetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridinamine

In a 8 mL vial5-nitro-2-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)pyridine(94.6 mg, 0.300 mmol) was dissolved in Tetrahydrofuran (THF) (3 mL) togive a pale yellow solution. Iron (84 mg, 1.498 mmol) and ammoniumchloride (80 mg, 1.498 mmol) were added followed by Water (1.500 mL).The reaction mixture was stirred at room temperature overnight.Additional iron (44 mg, 0.75 mmol) and ammonium chloride (40 mg, 0.75mmol) were added and the reaction mixture was stirred at roomtemperature for 6 hours. The reaction mixture was quenched with 10 mL ofan aqueous saturated solution of sodium bicarbonate and diluted with 25mL of EtOAc. The reaction mixture was filtered over a celite pad. Phaseswere separated by a separating funnel. The aqueous phase was extractedwith DCM (3×10 mL). The combined organic layers were evaporated and theresidue was purified by flash chromatography on silica gel using acolumn SNAP 25 g and Cyclohexane/ethyl acetate from 3:1 to 1:1 aseluents affording the title compound (82.9 mg) as colorless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 7.76 (1H, d), 7.09 (1H, dd), 7.02 (1H, t),6.75 (1H, d), 6.66 (1H, dd), 6.62 (1H, dd), 4.32 (1H, dd), 3.97 (1H,dd), 3.55 (2H, br. s.), 2.16-2.27 (1H, m), 1.61-1.76 (1H, m), 0.98-1.07(2H, m). UPLC_B: 0.78 min, 255 [M+H]+.

Intermediate 227 tert-butylN-[(1R)-1-[[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridyl]carbamoyl]-1-methyl-propyl]carbamate

In a 8 mL vial tube6-(1,1a,2,7btetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridinamine(Intermediate 226, 84 mg, 0.0305 mmol),(2R)-2-(tert-butoxycarbonylamino)-2-methyl-butanoic acid (59.7 mg, 0.275mmol) and DIPEA (0.080 mL, 0.458 mmol) were dissolved inN,N-Dimethylformamide (DMF) (2 mL) to give a pale yellow solution. HATU(151 mg, 0.397 mmol) was added. The reaction mixture was stirred at roomtemperature over week-end. The reaction mixture was evaporated in vacuoand the residue was purified by flash chromatography on silica gel usinga column SNAP 25 g and cyclohexane/ethyl acetate from 3:1 to 1:2 aseluents affording the title compound (74 mg) as white solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.14-8.18 (1H, m), 8.09-8.14 (1H, m), 7.04(1H, t), 6.85 (1H, d), 6.64-6.72 (2H, m), 6.30 (1H, br. s.), 4.91 (1H,br. s.), 4.31 (1H, dd), 3.94 (1H, dd), 1.83-2.17 (3H, m), 1.70-1.83 (1H,m), 1.51 (3H, s), 1.45 (9H, s), 0.91-1.05 (5H, m). UPLC_ipqc: 1.17 min,454 [M+H]+.

Intermediate 228(2R)—N-[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridyl]-2-amino-2-methyl-butanamide

In a 50 mL round-bottomed flask tert-butylN-[(1R)-1-[[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridyl]carbamoyl]-1-methyl-propyl]carbamate(Intermediate 227, 74 mg, 0.139 mmol) was dissolved in Dichloromethane(3 mL) to give a pale yellow solution. The reaction mixture was cooledat 0° C. and TFA (2 mL, 26.0 mmol) was added. The reaction mixture wasstirred at 0° C. for 2 hours. The reaction mixture was evaporated invacuo to give the crude product as a yellow oil. The sample was chargedon a 2 g SCX cartridge. It was then flushed with 36 mL of MeOH followedby 25 mL of 2M solution of ammonia in MeOH. The ammonia eluate wasevaporated in vacuo affording the title compound (46.2 mg) as a paleyellow oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 9.89 (1H, br. s.), 8.17-8.28 (2H, m), 7.04(1H, t), 6.81-6.88 (1H, m), 6.65-6.71 (2H, m), 4.31 (1H, dd), 3.95 (1H,dd), 2.06-2.14 (1H, m), 1.91-2.05 (1H, m), 1.53-1.77 (7H, m), 0.94 (3H,t), 0.90-1.05 (2H, m). LCMS UPLC/MS (method: IPQC2): rt=0.72 mins,MH+=354. UPLC_ipqc: 0.72 min, 354 [M+H]+.

Example 1(5R)-3-[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]-5-methyl-2,4-imidazolidinedione

A solution ofN1-[4-(1,3-dihydro-2-benzofuran-4-yloxy)phenyl]-D-alaninamide(Intermediate 7, 80 mg) and TEA (0.187 ml, 1.341 mmol) indichloromethane (10 ml) was stirred under argon at 0° C. Triphosgene(39.8 mg, 0.134 mmol) in dichloromethane (4 ml) was added and themixture was left under stirring at 0° C. for 45 minutes. An aqueoussaturated solution of NaHCO₃ was then added. The phases were separatedand the aqueous one was extracted 3 times with dichloromethane. Thegathered organic phases were dried over sodium sulphate and concentratedunder vacuum. The crude was purified by flash chromatography(FlashMasterPersonal), using as eluents a gradient Cyclohexane/Ethylacetate from 100:0 to 40:60. This afforded the title compound as a whitesolid (58.5 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.49-8.44 (1H, m), 7.39-7.32 (3H, m),7.18-7.13 (1H, m), 7.11-7.06 (2H, m), 6.91-6.87 (1H, m), 5.09-5.03 (2H,m), 4.91-4.87 (2H, m), 4.29-4.22 (1H, m), 1.36 (3H, d); UPLC-MS: 0.69min, 325 [M+1]+

Example 2(5R)-5-methyl-3-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-2,4-imidazolidinedione

N¹-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-D-alaninamide(Intermediate 14, 337 mg) was dissolved in 8.0 ml of ethyl acetate.Triethylamine (0.33 ml, 2.38 mmol) was added followed by a solution oftriphosgene (161 mg, 0.54 mmol) in 2.0 ml of ethyl acetate. After 5minutes stirring, DMAP (66 mg, 0.54 mmol) was added and the reactionmixture was stirred at room temperature for 10 minutes. After quenchingwith a saturated aqueous solution of NaHCO₃, the reaction mixture wasextracted two times with ethyl acetate. The gathered organic layers weredried over sodium sulphate, filtered and evaporated. The residue waspurified by silica gel chromatography eluting with a gradientCy-Hex/EtOAc from 100/0% to 0/100%. This afforded the title compound(160 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.49 (1H, br. s), 7.64-7.58 (1H, m),7.50-7.40 (3H, m), 7.34-7.25 (2H, m), 6.76 (1H, d), 4.34-4.18 (1H, m),2.58 (3H, s), 1.38 (3H, d); UPLC-MS_B: 0.73 min, 338 [M+H]+.

Example 3(5R)-3-{4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione

N¹-{4-[(3,6-dimethyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-D-alaninamide(Intermediate 23, 54 mg) was dissolved in 7.0 ml of ethyl acetate. Thentriethylamine (0.051 ml, 0.37 mmol) was added followed by a solution oftriphosgene (24.6 mg, 0.083 mmol) in 2.0 ml of ethyl acetate. Afterstirring for 5 minutes, DMAP (10.1 mg, 0.083 mmol) was added and thereaction mixture was stirred at room temperature for 10 minutes. Afterquenching with an aqueous solution of NaHCO₃, the mixture was extractedtwo times with ethyl acetate and the collected organic layers were driedover sodium sulphate, filtered and evaporated. The residue was purifiedby silica gel chromatography eluting with a gradient c-Hex/EtOAc from100/0 to 0/100 to afford the title compound (11 mg)

¹H-NMR (400 MHz, CD₃OD): δ ppm 7.50-7.45 (2H, m), 7.26-7.16 (3H, m),6.63-6.57 (1H, m), 4.36-4.26 (1H, m), 2.57 (3H, s), 2.45 (3H, s), 1.52(3H, d); UPLC-MS_B: 0.78 min, 352 [M+H]+.

Example 45,5-dimethyl-3-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}-2,4-imidazolidinedione

2-methyl-N¹-{4-[(3-methyl-1,2-benzisoxazol-4-yl)oxy]phenyl}alaninamide(Intermediate 25, 18 mg) was dissolved in 4 ml of ethyl acetate.Triethylamine (0.017 ml, 0.12 mmol) was then added followed by asolution of triphosgene (8.21 mg, 0.028 mmol) in 1.0 ml of ethylacetate. After stirring for 5 minutes, DMAP (3.4 mg, 0.028 mmol) wasadded and the reaction mixture was stirred at room temperature for 10minutes. After quenching with a saturated aqueous solution of NaHCO₃,the mixture was extracted two times with ethyl acetate and the collectedorganic layers were dried over sodium sulphate, filtered and evaporated.The crude obtained was charged on a silica gel column and eluted withc-Hex/EtOAc as eluents (from all 100:0 to 0:100). This afforded 10 mg ofthe title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.57 (1H, br. s), 7.66-7.57 (1H, m),7.51-7.43 (3H, m), 7.27 (2H, d), 6.76 (1H, d), 2.57 (3H, s), 1.43 (6H,s); UPLC-MS: 0.72 min, 352 [M+H]+.

Example 5(5R)-5-ethyl-3-{6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione

(2R)-2-amino-N-{6-[(3-ethyl-1,2-benzisoxazol-4-yl)oxy]-3-pyridinyl}butanamide(Intermediate 35) was dissolved in dichloromethane (1.0 ml) and TEA(0.004 ml, 0.03 mmol) was added. The reaction mixture was cooled down to0° C. and triphosgene (1.3 mg, 4.49 μmol) dissolved in 0.1 ml ofdichloromethane was added. The reaction mixture was stirred at thattemperature for 30 minutes. The mixture was quenched with 0.5 ml ofwater and water was removed by addition of sodium sulphate. The organicphase was pipetted off and evaporated and the crude obtained was chargedon a silica gel column (Biotage SP1 system) and eluted withCyhexane/EtOAv (from 1/0 to 7/3, then 7/3, then from 7/3 to 1/1, the1/1, then from 1/1 to 0/1) to afford the title compound (1.3 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.35-8.31 (1H, m), 7.92-7.87 (1H, m),7.60-7.52 (1H, m), 7.46-7.41 (1H, m), 7.19-7.13 (1H, m), 7.05-7.00 (1H,m), 5.57 (1H, br. s), 4.29-4.23 (1H, m), 2.94 (2H, q), 2.12-1.91 (2H,m), 1.39 (3H, t), 1.11 (3H, t). UPLC: 0.68 min, 367 [M+H]+.

Example 6(5R)-5-ethyl-3-(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

(2R)-2-amino-N-(6-{[3-(1-methylethyl)-1,2-benzisoxazol-4-yl]oxy}-3-pyridinyl)butanamide(Intermediate 44) was dissolved in dichloromethane (1.0 ml) and TEA (3μL, 0.022 mmol) was added. The reaction mixture was cooled down to 0° C.and triphosgene (0.6 mg, 1.98 μmol) was added and the mixture wasstirred at that temperature for 30 minutes. The reaction mixture wasquenched with water and water was removed by addition of sodiumsulphate. The organic phase was pipetted off and evaporated and theresidue obtained was charged on a silica gel column (Biotage SP1 system)and eluted with Cyhexane/EtOAc (from all 1/0 to 7/3, then 7/3, then from7/3 to 1/1, then 1/1, then 0/1) to afford the title compound (1 mg).

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.39-8.34 (1H, m), 7.93-7.87 (1H, m),7.59-7.51 (1H, m), 7.46-7.38 (1H, m), 7.20-7.14 (1H, m), 7.06-6.97 (1H,m), 5.50 (1H, br. s), 4.31-4.23 (1H, m), 3.42-3.32 (1H, m), 2.08-1.91(2H, m), 1.45 (6H, d), 1.11 (3H, t). UPLC: 0.72 min, 381 [M+H]+.

Example 7(5R)-3-{4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}-5-methyl-2,4-imidazolidinedione

To a solution ofN¹-{4-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]phenyl}-D-alaninamide(Intermediate 57, 66 mg) in dichloromethane (20 ml, SCRC) was addedtriethylamine (0.085 ml, 0.607 mmol, SCRC). Triphosgene (24.00 mg, 0.081mmol, SCRC) was added dropwise at 0° C. and the reaction mixture wasstirred at room temperature for 0.5 hours. The mixture was quenched withwater (20 ml) and it was extracted with dichloromethane (3 times 50 ml,SCRC). The combined organic layers were dried, evaporated and purifiedby column chromatography on silica gel with EtAc/PE (1/30) as eluents toafford the title compound as a white solid (40 mg).

¹H NMR (CDCl₃): δ ppm 7.36-7.34 (2H, m), 7.09-7.03 (3H, m), 6.60-6.58(1H, d), 6.41-6.39 (1H, d), 5.60 (1H, s), 4.27-4.25 (3H, m), 1.58-1.51(3H, d), 1.42 (6H, s); MS_(—)2 (ESI): 353 [M+H]+

Example 8(5R)-3-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-methyl-2,4-imidazolidinedione

N¹-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-D-alaninamide(Intermediate 61, 28 mg) was dissolved in dry dichloromethane (3 ml).The reaction mixture was cooled down in an ice bath. Triethylamine (71.5μl, 0.513 mmol) was added. Then a solution of triphosgene in drydichloromethane (11.42 mg, 0.038 mmol dissolved in 1 ml ofdichloromethane) was added dropwise.

The reaction mixture was stirred at 0° C., under argon, during 15 min. Asaturated aqueous solution of NaHCO₃ was added (4 ml) and the aqueouslayer was extracted with dichloromethane 4 times (4×5 ml). After dryingover sodium sulphate, the solvents were removed under vacuum. Theresidue obtained was purified by silica gel chromatography (Companionsystem, 2×4 g silica cartridges) with a gradientcyclohexane/ethylacetate from 100/0 to 50/50. This afforded the titlecompound as a film (17.5 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.31 (1H, d), 7.77 (1H, dd), 7.13 (1H,t), 7.00 (1H, d), 6.66 (1H, d), 6.56 (1H, d), 5.81 (1H, br.s), 4.29 (1H,dd), 4.23 (2H, s), 1.58 (3H, d), 1.37 (6H, s); UPLC_B: 0.76 min, 354[M+H]+

Example 9(5R)-3-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-2,4-imidazolidinedione

The title compound was made in a similar fashion to the preparation ofExample 8 replacingN¹-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-D-alaninamide(Intermediate 61) with(2R)-2-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide(Intermediate 63, 57 mg). This afforded the title compound as a whitesolid (40 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.30 (1H, d), 7.77 (1H, dd), 7.13 (1H,t), 7.01 (1H, d), 6.67 (1H, d), 6.57 (1H, d), 5.96 (1H, br.s), 4.23 (2H,s), 4.20 (1H, m), 2.03-1.97 (1H, m), 1.93-1.87 (1H, m), 1.38 (6H, s),1.06 (3H, t); UPLC: 0.73 min, 368 [M+H]+

Example 10(5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-ethyl-2,4-imidazolidinedione

To a solution of(2R)-2-amino-N-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}butanamide(Intermediate 67, 3 mg) in dry dichloromethane (0.5 ml), TEA (6.11 μL,0.044 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (1.170 mg, 3.94 μmol) in dry dichloromethane(0.125 ml) was then added dropwise and the reaction mixture was stirredat the 0° C. for 30 minutes. The reaction was quenched with water (3ml), and the organic phase was separated, dried over sodium sulphate,filtered and evaporated. The residue was purified by flashchromatography on silica gel using a column Isolute (1 g) anddichloromethane/methanol from 99.5:0.5 to 9:10 as eluent to afford thetitle compound (0.7 mg) as a white solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 8.73 (2H, s), 7.18 (1H, t), 6.72 (1H, d),6.63 (1H, d), 5.58 (1H, bs), 4.28-4.23 (1H, m), 4.24 (2H, s), 2.09-1.89(2H, m), 1.37 (6H, s). 1.09 (3H, t).

Example 117-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5,7-diazaspiro[3.4]octane-6,8-dione

1-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}cyclobutanecarboxamide(Intermediate 70, 17 mg) was dissolved in dry dichloromethane (1.8 ml).The reaction mixture was cooled down in an ice bath. Triethylamine(39.48 μl, 0.283 mmol) was added at 0° C. Then 0.89 ml of a solution oftriphosgene in dry dichloromethane was added dropwise (0.0135 mg, 4.00mg). The reaction mixture was stirred under argon during 10 min at 0°C., then during 30 min at room temperature. Then an additional 0.25equivalent of triphosgene in dichloromethane (0.26 M solution) was addedat 0° C. and the reaction mixture was stirred under argon an additional30 min at room temperature. The solvents were removed under vacuum. Theresidue obtained was purified by flash chromatography on silica gel(Companion system, 4 g silica cartridge) with cyclohexane/ethylacetateas eluents from 100/0 to 55/45. This afforded the title compound (12mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.31 (1H, d), 7.76 (1H, dd), 7.13 (1H,t), 7.11 (1H, d), 6.67 (1H, d), 6.65 (1H, d), 5.70 (1H, s), 2.50 (2H,s), 4.74 (2H, m), 2.43 (2H, m), 2.24 (1H, m), 1.94 (1H, m), 1.38 (6H,m); UPLC: 0.72 min, 380 [M+H]+

Example 126-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-4,6-diazaspiro[2.4]heptane-5,7-dione

The title compound (5 mg, 49% yield) was made in a similar fashion tothe preparation of Example 11 replacing1-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}cyclobutanecarboxamide(Intermediate 70) with1-amino-N-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}cyclopropanecarboxamide(Intermediate 73, 9 mg).

¹H NMR (400 MHz, MeOD): δ ppm 8.24 (1H, m), 7.93 (1H, m), 7.14 (1H, t),7.07 (1H, d), 6.64 (1H, d), 6.54 (1H, d), 4.22 (2H, s), 1.51 (2H, m),1.42 (2H, m), 1.35 (6H, s); UPLC: 0.78 min, 366 [M+H]+

Example 133-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione

N1-{6-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-2-methylalaninamide(Intermediate 75, 7.5 mg) was dissolved in dry dichloromethane (1 ml).The reaction mixture was cooled down in an ice bath. Triethylamine(18.37 μl, 0.132 mmol) was added at 0° C. Then 0.5 ml of a solution oftriphosgene in dry dichloromethane (0.012 mmol) was added dropwise. Thesolution was prepared with 7.18 mg of triphosgene dissolved in 1 ml ofdichloromethane). The reaction mixture was stirred under argon during 10min at 0° C., then during 30 min at room temperature. A saturatedaqueous solution of NaHCO₃ was added and the aqueous layer was extractedwith dichloromethane 4 times. After drying over sodium sulphate, thesolvents were removed under vacuum.

The residue obtained was purified by flash chromatography on silica gel(Companion system, 4 g silica cartridge) with a gradientcyclohexane/ethylacetate from 100/0 to 55/45. This afforded the titlecompound (4.5 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.31 (1H, d), 7.77 (1H, dd), 7.12 (1H,t), 6.99 (1H, d), 6.66 (1H, d), 6.56 (1H, d), 5.47 (1H, br s), 4.22 (2H,s), 1.56 (6H, s), 1.36 (6H, s); UPLC: 0.70 min, 368 [M+H]+

Example 14(5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-(1,1-dimethylethyl)-2,4-imidazolidinedione

N1-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-3-methyl-D-valinamide(Intermediate 78, 9.9 mg) was dissolved in dry dichloromethane (1 ml).The reaction mixture was cooled down in an ice bath. Triethylamine(22.35 μl, 0.160 mmol) was added. The reaction mixture was cooled downat 0° C. Then 0.5 ml of a solution of triphosgene in dry dichloromethane(0.015 mmol) was added dropwise. The reaction mixture was stirred underargon during 20 min at 0° C. Some water (2 ml) was added and the aqueouslayer was extracted with dichloromethane 4 times. After drying oversodium sulphate, the solvents were removed under vacuum. The residueobtained was purified by flash chromatography on silica gel (Companionsystem, 4 g silica cartridge) with cyclohexane/ethylacetate as eluentsfrom 100/0 to 60/40. This afforded the title compound (7.7 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.68 (2H, s), 7.17 (1H, t), 6.71 (1H, d),6.62 (1H, d), 6.19 (1H, s), 4.23 (2H, s) 3.92 (1H, s) 1.36 (6H, s) 1.12(9H, s); UPLC: 0.75 min, 397 [M+H]+

Example 15(5R)-5-ethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution of(2R)-2-amino-N-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]butanamide(Intermediate 89, 90 mg) in dry dichloromethane (15 ml) TEA (0.185 ml,1.326 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (35.4 mg, 0.119 mmol) in dry dichloromethane (5mL) was slowly added and the reaction mixture was stirred for 30 minutesat the same temperature. The reaction was quenched with water (10 ml)and two phases were separated. The organic layer was dried over sodiumsulphate, filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 8:2 to cyclohexane/ethyl acetate 1:1 aseluents affording the title compound (65 mg, 0.178 mmol) as a whitesolid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.63 (1H, s), 8.14 (1H, d), 7.85 (1H,dd), 7.11 (1H, s), 7.09 (1H, t), 6.68 (1H, dd), 6.52 (1H, dd), 4.45 (2H,s), 4.18-1.24 (1H, m), 1.76-1.88 (1H, m), 1.64-1.76 (1H, m), 1.13-1.18(2H, m), 0.96 (3H, t), 0.89-0.94 (2H, m); UPLC_B: 0.78 min, 366 [M+H]+.

Example 165,5-dimethyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution of2-methyl-N1-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]alaninamide(Intermediate 91, 34 mg) in dry dichloromethane (6 mL) TEA (0.070 mL,0.501 mmol) was added and the mixture was cooled to 0° C. A solution oftriphosgene (13.38 mg, 0.045 mmol) in dry dichloromethane (2 mL) wasslowly added and the reaction mixture was stirred for 1 hour at the sametemperature. The reaction was quenched with water (3 ml) and two phaseswere separated. The organic layer was dried over sodium sulphate,filtered and evaporated. The residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnSNAP and cyclohexane/ethyl acetate 7:3 to cyclohexane/ethyl acetate 3:7as eluents affording the title as a white solid (23 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.63 (1H, s), 8.17 (1H, d), 7.88 (1H,d), 7.06-7.12 (2H, m), 6.67 (1H, d), 6.51 (1H, d), 4.45 (2H, s), 1.41(6H, s), 1.12-1.17 (2H, m), 0.88-0.93 (2H, m); UPLC: 0.73 min, 366[M+H]+.

Example 17(5R)-5-ethyl-5-methyl-3-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution ofN1-[6-(spiro[1-benzofuran-3,1′-cyclopropan]-4-yloxy)-3-pyridinyl]-D-isovalinamide(Intermediate 93, 68 mg) and TEA (0.134 mL, 0.962 mmol) in drydichloromethane (11 ml) at 0° C. was added dropwise a solution oftriphosgene (25.7 mg, 0.087 mmol) in dry dichloromethane (3.14 m) andthe mixture thus obtained was stirred at the same temperature. After 1hour a solution of triphosgene (25.7 mg, 0.087 mmol) in drydichloromethane (3.14 ml) was added. After 3 hours UPLC/MS showed theabsence of the starting material and the presence of the desiredcompound. Water was then added, the organic phase was separated and theaqueous one was extracted again with dichloromethane. The organic phasewas washed with brine, dried over sodium sulphate and concentrated undervacuum to give 87 mg of crude. This was purified by flash chromatography(Biotage KP-Sil 10 g SNAP column, eluant cyclohexane/ethyl acetate from88/12 to 0/100 in 10CV) to give 47 mg of the title compound as a whitefoam.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.59 (1H, s), 8.14 (1H, d), 7.85 (1H,dd), 7.08 (2H, t), 6.66 (1H, d), 6.50 (1H, d), 4.44 (2H, s), 1.70-1.84(1H, m), 1.57-1.70 (1H, m), 1.38 (3H, s), 1.08-1.19 (2H, m), 0.89-0.95(2H, m), 0.85 (3H, t); UPLC: 1.04 min, 380 [M+H]+.

Example 18(5R)-5-ethyl-3-(6-{[(3S/R)-3-methyl-1,3-dihydro-2-benzofuran-4-yl]oxy}-3-pyridinyl)-2,4-imidazolidinedione(Diastereoisomeric Mixture)

In a 50 ml round-bottomed flask(2R)-2-amino-N-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide(Intermediate 102, 24.4 mg) was dissolved in dichloromethane (3 ml) togive a pale yellow solution which was cooled at 0° C. TEA (0.049 ml,0.354 mmol) was added. A solution of triphosgene (9.46 mg, 0.032 mmol)in 0.7 ml of dichloromethane was added dropwise to the reaction mixtureat 0° C. After 20 minutes, the reaction mixture was quenched with 5 mlof water and diluted with 5 ml of dichloromethane. Phases were separatedthrough a phase separator cartridge. The organic phase was evaporatedunder vacuum to afford the crude product which was purified by silicagel chromatography (Biotage SP1 system, 10 g SNAP Silica column) withCyclohexane/EtOAc as eluents (from 2/1 to 1/2 in 15 CV; then 1/2 for 10CV). The collected fractions afforded the title compound as a 1:1mixture of diastereoisomers (20.2 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.27 (1H, d), 7.78 (1H, dd), 7.33 (1H,t), 7.09 (1H, d), 7.06-6.96 (2H, m), 5.91 (1H, br. s.), 5.40-5.29 (1H,m), 5.20 (1H, dd), 5.09 (1H, d), 4.25-4.18 (1H, m), 2.08-1.84 (2H, m),1.45 (3H, d), 1.07 (3H, t). UPLC_ipqc: 0.87 min, 354 [M+H]+

Example 19 and Example 20(5R)-5-ethyl-3-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Diastereoisomers 1 and 2)

Both diastereoisomers of Example 18 were separated by preparative chiralchromatography.

Chiral preparative HPLC conditions were the following ones: Column:Chiralpak AS-H (25×2 cm), 5u; Mobile phase: n-Hexane/Ethanol 70:30% v/v;Flow rate: 15 ml/min; UV: 220 nm; Sample preparation: 20 mg dissolved in1 ml of hexane/ethanol 1:1 v/v; Sample concentration: 20 mg/ml;Injection volume: 1000 uL.

Chiral Analytical Chromatography conditions were the following ones:Column: Chiralpak AS-H (25×0.46 cm); Mobile phase: n-Hexane/Ethanol70:30% v/v; Flow rate: 0.8 ml/min; DAD: 210-340 nm; CD: 240 nm.

This chiral preparative HPLC afforded

Example 19 which was diastereoisomer 1 of(5R)-5-ethyl-3-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(7.1 mg). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.27 (d, 1H), 7.78 (dd, 1H),7.33 (t, 1H), 7.09 (d, 1H), 6.96-7.06 (m, 2H), 5.91 (br. s., 1H),5.29-5.40 (m, 1H), 5.20 (dd, 1H), 5.09 (d, 1H), 4.18-4.25 (m, 1H),1.84-2.08 (m, 2H), 1.45 (d, 3H), 1.07 (t, 3H). UPLC-MS_ipqc: 0.87 min,354 [M+1]+. Chiral Analytical Chromatography HPLC: r.t.=12.327 min, 100%d.e.

Example 20 which was diastereoisomer 2 of(5R)-5-ethyl-3-{6-[(3-methyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(8.0 mg). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.27 (d, 1H), 7.78 (dd, 1H),7.33 (t, 1H), 7.09 (d, 1H), 6.96-7.06 (m, 2H), 5.91 (br. s., 1H),5.29-5.40 (m, 1H), 5.20 (dd, 1H), 5.09 (d, 1H), 4.18-4.25 (m, 1H),1.84-2.08 (m, 2H), 1.45 (d, 3H), 1.07 (t, 3H). UPLC-MS_ipqc: 0.87 min,354 [M+1]+. Chiral Analytical Chromatography HPLC: r.t.=16.579 min, 100%d.e.

Example 21(5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Diastereoisomeric Mixture)

In a 50 ml round-bottomed flask(2R)-2-amino-N-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide(Intermediate 110, 9 mg) was dissolved in dichloromethane (2 ml) to givea pale yellow solution that was cooled at 0° C. TEA (0.017 ml, 0.119mmol) was added. 0.5 ml of a solution of triphosgene (14 mg in 2 ml ofdichloromethane) was added dropwise to the reaction mixture at 0° C. Thereaction mixture was stirred at 0° C. After 20 minutes, the reaction wascompleted. The reaction mixture was evaporated under vacuum to affordthe crude product as a pale yellow oil which was purified via BiotageSP1 (with Cyclohexane/EtOAc as eluents from 2:1 to 1:2 in 15 CV; then1:2 for 10 CV; 10 g SNAP Silica column). The collected fractionsafforded the title compound(5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(6.9 mg) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.29 (1H, d), 7.80 (1H, dd), 7.35 (1H, t),7.11 (1H, d), 6.99-7.07 (2H, m), 5.91 (1H, br. s.), 5.25-5.33 (1H, m),5.21 (1H, dd), 5.14 (1H, d), 4.21-4.26 (1H, m), 1.82-2.13 (3H, m),1.69-1.81 (1H, m), 1.09 (3H, t), 0.93 (3H, t). UPLC_B: 0.78 min, 368[M+H]+.

Example 22 and Example 23(5R)-5-ethyl-3-{6[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Diastereoisomers 1 and 2)

Both diastereoisomers of Example 21 were separated by preparative chiralchromatography affording two fractions.

Chiral preparative HPLC conditions were the following ones: ColumnChiralpak AD-H (25×2 cm) 5 μm; Mobile phase n-Hexane/2-Propanol 85:15%v/v; Flow rate (ml/min) 15; UV detection 220 nm; Sample preparation 4 mgdissolved in 2 ml of methanol/ethanol 50:50% v/v; Sample concentration 2mg/ml; Injection volume 2000 μl (equivalent to 4 mg)

Chiral Analytical Chromatography conditions were the following ones:Column: Chiralpak AD-H (25×0.46 cm); Mobile phase: n-Hexane/2-Propanol85:15% v/v; Flow rate: 0.8 ml/min; DAD: 210-340 nm; CD: −.

This chiral preparative HPLC afforded:

Example 22 which was diastereoisomer 1 of(5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(2.3 mg). ¹H NMR (400 MHz, MeOD) δ ppm 8.17 (1H, d), 7.89 (1H, dd), 7.37(1H, t), 7.17 (1H, d), 7.10 (1H, d), 7.01 (1H, d), 5.13-5.23 (2H, m),5.08 (1H, d), 4.25 (1H, dd), 1.78-2.01 (3H, m), 1.64-1.78 (1H, m), 1.05(3H, t), 0.81-0.89 (3H, m). NH missed. UPLC_B: 0.76 min, 368 [M+1]+.Chiral Analytical Chromatography HPLC: 14.93 min, 98.6% d.e.

Example 23 which was diastereoisomer 2 of(5R)-5-ethyl-3-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(2.6 mg). ¹H NMR (400 MHz, MeOD) δ ppm 8.18 (1H, d), 7.89 (1H, dd), 7.38(1H, t), 7.17 (1H, d), 7.10 (1H, d), 7.02 (1H, d), 5.13-5.24 (2H, m),5.09 (1H, d), 4.25 (1H, dd), 1.79-2.05 (3H, m), 1.65-1.78 (1H, m), 1.06(3H, t), 0.85 (3H, t). NH missed. UPLC_B: 0.76 min, 368 [M+1]+. ChiralAnalytical Chromatography HPLC: 17.51 min, 100% d.e.

Example 245,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Racemic Mixture)

In a 50 ml round-bottomed flask2-methyl-N1-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}alaninamide(Intermediate 121, 72.6 mg) was dissolved in dichloromethane (5 ml) togive a pale yellow solution. TEA (0.142 ml, 1.021 mmol) was added andthe reaction mixture was cooled at 0° C. A solution of triphosgene (27.3mg, 0.092 mmol) in 1 ml of dichloromethane was added to the reactionmixture which was stirred at 0° C. After 15 minutes, additional solutionof triphosgene (27.3 mg, 0.092 mmol) in 1 ml of dichloromethane wasadded to the reaction mixture. After 15 minutes, the reaction mixturewas evaporated under vacuum to give the crude product which was purifiedvia Biotage SP1 (using Cyclohexane/EtOAc as eluents from 3:1 to 1:2 in10 CV; then 1:2 for 5 CV; 10 g SNAP Silica column). The collectedfractions afforded the title compound5,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedioneas a colourless oil (41.6 mg).

¹H NMR (400 MHz, CDCl₃) δ ppm 8.28 (1H, d), 7.75 (1H, dd), 7.13 (1H, t),6.95 (1H, d), 6.74 (1H, dd), 6.65 (1H, dd), 5.76 (1H, br. s.), 4.15-4.20(1H, m), 3.63-3.74 (1H, m), 2.72-2.82 (1H, m), 2.06-2.23 (2H, m), 1.57(6H, s), 1.02 (3H, d). UPLC_B: 0.80 min, 368 [M+H]+.

Example 25 and Example 265,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Enantiomers 1 and Enantiomer 2)

Both enantiomers of Example 24 were separated by preparative chiralchromatography.

Chiral preparative HPLC conditions were the following ones: Column:Chiralpak AD-H (25×2 cm), 5u; Mobile phase: n-Hexane/Ethanol 40/60 v/v;Flow rate: 15 ml/min; UV: 220 nm; Sample preparation: 50 mg dissolved in2 ml of ethanol Added 1 ml n-Hexane; Sample concentration: 16.7 mg/ml;Injection volume: 1000 μL.

Chiral analytical chromatography conditions were the following ones:Column: Chiralpak AD-H (25×0.46 cm); Mobile phase: n-Hexane/Ethanol40:60% v/v; Flow rate: 0.8 ml/min; DAD: 210-340 nm.

This preparative chiral chromatography afforded

Example 25 which was the enantiomer 1 of5,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(16.6 mg). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.28 (1H, d), 7.75 (1H, dd),7.13 (1H, t), 6.95 (1H, d), 6.74 (1H, dd), 6.65 (1H, dd), 5.76 (1H, br.s.), 4.15-4.20 (1H, m), 3.63-3.74 (1H, m), 2.72-2.82 (1H, m), 2.06-2.23(2H, m), 1.57 (6H, s), 1.02 (3H, d). UPLC_B: 0.81 min, 368 [M+H]+.Chiral Analytical Chromatography HPLC: 12.48 min, 100.0% e.e.

Example 26 which was the enantiomer 2 of5,5-dimethyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(17.0 mg). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.28 (1H, d), 7.75 (1H, dd),7.13 (1H, t), 6.95 (1H, d), 6.74 (1H, dd), 6.65 (1H, dd), 5.76 (1H, br.s.), 4.15-4.20 (1H, m), 3.63-3.74 (1H, m), 2.72-2.82 (1H, m), 2.06-2.23(2H, m), 1.57 (6H, s), 1.02 (3H, d). UPLC_B: 0.81 min, 368 [M+H]+.Chiral Analytical Chromatography HPLC: 14.68 min, 98.4% e.e.

Example 275,5-dimethyl-3-{6[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Racemic Mixture)

To a solution of2-methyl-N1-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}alaninamide(Intermediate 129, 100 mg) in dry dichloromethane (10 ml) TEA (0.177 ml,1.273 mmol) was added and the reaction mixture was cooled to 0° C. Atthis point a solution of triphosgene (37.8 mg, 0.127 mmol) in drydichloromethane (2.5 ml) was slowly added (over 30 minutes) and thereaction mixture was stirred for 1 hour at the same temperature. Thereaction was then quenched with water and the two phases were separated.The organic layer was dried over sodium sulphate, filtered andevaporated and the residue was purified by silica gel chromatography(Companion system, with a gradient from cyclohexane/ethyl acetate 8:2 tocyclohexane/ethyl acetate 1:1) to give the title compound as a whitesolid (40 mg, 0.103 mmol).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.33 (1H, d), 7.77 (1H, dd), 7.07 (1H,t), 6.97 (1H, d), 6.74 (2H, dd), 5.60 (1H, br. s.), 4.19 (1H, d), 3.72(1H, d), 1.88 (1H, dd), 1.59 (6H, s), 1.18-1.26 (4H, m), 0.87 (1H, dd);UPLC_ipqc: 1.03 min, 380 [M+H]+.

Example 28 and Example 295,5-dimethyl-3-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Enantiomer 1 and Enantiomer 2)

Both enantiomers of Example 27 were separated by semi-preparative chiralSFC chromatography. Chiral SFC conditions were the following ones:Column: Chiralpak IC (25×2.1 cm); Mobile phase: Ethanol+0.1%I-propylamine 20%; Flow rate: 45 ml/min; Pressure: 120 bar; UV: 220 nm;

This semi-preparative chiral SFC chromatography on 30 mg of Example 27afforded:

Example 28 which was the enantiomer 1 of5,5-dimethyl-3-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(12 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.33 (1H, d), 7.77 (1H, dd), 7.07 (1H,t), 6.97 (1H, d), 6.74 (2H, dd), 5.81 (1H, s), 4.19 (1H, d), 3.72 (1H,d), 1.88 (1H, dd), 1.60 (6H, s), 1.16-1.24 (4H, m), 0.87 (1H, dd);Chiral Analytical SFC Chromatography: 7.51 min, 100.0% e.e.

and Example 29 which was the enantiomer 2 of5,5-dimethyl-3-{6-[(1a-methyl-1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(14 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.33 (1H, d), 7.77 (1H, dd), 7.07 (1H,t), 6.97 (1H, d), 6.74 (2H, dd), 5.87 (1H, s), 4.19 (1H, d), 3.72 (1H,d), 1.88 (1H, dd), 1.59 (6H, s), 1.19-1.23 (4H, m), 0.87 (1H, dd);Chiral Analytical SFC Chromatography: 9.40 min, 100.0% e.e.

Example 30(5R)-5-ethyl-5-methyl-3-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution ofN1-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-isovalinamide(Intermediate 141, 40 mg) in dry dichloromethane (8 mL) TEA (0.076 mL,0.544 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (14.54 mg, 0.049 mmol) in dry dichloromethane (4ml) was slowly added and the reaction mixture was stirred for 3 hours atthe same temperature. The reaction was quenched with water (10 ml) andtwo phases separated. The organic layer was dried over sodium sulphate,filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using 10 g SNAP column andcyclohexane/ethyl acetate as eluents from 7:3 to 3:7 as eluent. Thisafforded the title compound (25 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.60 (1H, s), 8.15 (1H, d), 7.87 (1H,dd), 7.15 (1H, t), 7.10 (1H, d), 6.95 (1H, d), 6.79 (1H, d), 4.85 (2H,s), 3.54 (2H, s), 1.72-1.84 (1H, m), 1.60-1.72 (1H, m), 1.39 (3H, s),1.33-1.38 (2H, m), 0.87 (3H, t), 0.62-0.67 (2H, m); UPLC_IPQC: 0.97 min,394 [M+H]+.

Example 313-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5,5-dimethyl-2,4-imidazolidinedione

To a solution ofN1-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-2-methylalaninamide(Intermediate 143, 18 mg, 0.053 mmol) in dry Dichloromethane (DCM) (3mL) TEA (0.037 ml, 0.263 mmol) was added and the reaction mixture wascooled to 0° C. A solution of triphosgene (7.02 mg, 0.024 mmol) in dryDichloromethane (1 mL) was slowly added and the reaction mixture wasstirred for 2 hours while the temperature was allowed to reach r.t. Thereaction was quenched with water (3 ml) and two phases were separated.The organic layer was dried (Na₂SO₄), filtered and evaporated and theresidue was purified by flash chromatography (Biotage system) on silicagel using a column SNAP 10 g and cyclohexane/ethyl acetate from 8:2 to1:1 as eluents affording the title compound (13 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.72-8.77 (3H, m), 7.17 (1H, t),6.65-6.75 (2H, m), 4.23 (2H, s), 1.43 (6H, s), 1.25 (6H, s). UPLC: 0.67min, 369 [M+H]+.

Example 32(5R)-3-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-5-(1-methylethyl)-2,4-imidazolidinedione

To a solution ofN1-{2-[(3,3-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-D-valinamide(Intermediate 145, 38 mg, 0.107 mmol) in dry Dichloromethane (DCM) (5mL) TEA (0.074 mL, 0.533 mmol) was added and the reaction mixture wascooled to 0° C. A solution of triphosgene (14.24 mg, 0.048 mmol) in dryDichloromethane (DCM) (1 mL) was slowly added and the reaction mixturewas stirred for 30 minutes at the same temperature. The reaction wasquenched with water (3 ml) and two phases were separated. The organiclayer was dried (Na₂SO₄), filtered and evaporated and the residue waspurified by flash chromatography (Biotage system) on silica gel using acolumn SNAP 25 g and cyclohexane/ethyl acetate from 8:2 to 1:1 as eluentaffording the title compound (15 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.77 (1H, br.s), 8.62 (2H, s), 7.18(1H, t), 6.65-6.75 (2H, m), 4.22 (2H, s), 4.15-4.20 (1H, m), 2.10-2.20(1H, m), 1.25 (6H, s), 1.03 (3H, d), 0.90 (3H, d). UPLC: 0.71 min, 383[M+H]+.

Example 33(5R)-3-{6-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-2,4-imidazolidinedione

((2R)-2-amino-N-{6-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide)(Intermediate 209, 10 mg, 0.029 mmol) was dissolved in drydichloromethane (1 ml). The reaction mixture was cooled down in an icebath. Triethylamine (0.024 ml, 0.176 mmol) was added at 0° C. Then 0.5ml of a solution of triphosgene in dry dichloromethane (4.78 mg, 0.016mmol in 0.5 ml) was added dropwise. The reaction mixture was stirredunder argon during 20 min at 0° C. A saturated aqueous solution ofNaHCO₃ was added (3 ml). The aqueous layer was extracted withdichloromethane 4 times (4×4 mL). Combined organic layers were driedover Na₂SO₄ and evaporated. the residue was purified by flashchromatography (Companion system) on silica gel using a 4 g silicacartridge and cyclohexane/ethyl acetate from 100:0 to 60:40 as eluentsaffording the title compound (7 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 8.29 (1H, d), 7.74 (1H, dd), 7.14 (1H,d), 7.00 (1H, d), 6.61 (1H, dd), 6.55 (1H, d), 5.91 (1H, br.s),4.20-4.25 (1H, m), 3.02 (2H, s), 1.85-2.10 (2H, m), 1.51 (6H, s), 1.08(3H, t). UPLC_B: 1.02 min, 368 [M+H]+.

Example 345,5-dimethyl-3-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]-2,4-imidazolidinedione

To a solution of2-methyl-N1-[6-(1H-spiro[2-benzopyran-4,1′-cyclopropan]-5-yloxy)-3-pyridinyl]alaninamide(34 mg, 0.096 mmol) in dry Dichloromethane (DCM) (5 mL) TEA (0.067 mL,0.481 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (12.85 mg, 0.043 mmol) in dry Dichloromethane(DCM) (2 mL) was slowly added and the reaction mixture was stirred for30 minutes at the same temperature. The reaction was quenched with water(5 ml) and two phases were separated. The organic layer was dried(Na₂SO₄), filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a column SNAP 10 gand Dichloromethane/methanol 99:1 to Dichloromethane/methanol 95:5 aseluents affording the title compound (35 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.63 (1H, s), 8.18 (1H, d), 7.90 (1H,dd), 7.16 (1H, t), 7.10 (1H, d), 6.96 (1H, dd), 6.80 (1H, dd), 4.86 (2H,s), 3.55 (2H, s), 1.43 (6H, s), 1.34-1.39 (2H, m), 0.63-0.68 (2H, m).UPLC_B: 0.92 min, 380 [M+H]+.

Example 35(5R)-3-[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]-5-ethyl-5-methyl-2,4-imidazolidinedione

To a solution ofN1-[2-(2,3-dihydrospiro[chromene-4,1′-cyclopropan]-5-yloxy)-5-pyrimidinyl]-D-isovalinamide(Intermediate 225, 4 mg, 10.86 μmol) in dry Dichloromethane (1 mL) TEA(3.78 μL, 0.027 mmol) was added. The reaction was cooled in an ice-bath,then a solution of triphosgene (1.450 mg, 4.89 μmol) in dryDichloromethane (0.250 mL) was added once then two other times with 10minutes in between. The reaction was stirred at 0° C. for 40 minutes,then it was quenched with water (5 mL) maintaining the reaction in theice-bath. The organic layer was separated, dried (Na₂SO₄), filtered andevaporated. The residue was purified by chromatography on silica gelusing a column isolute 2 g and cyclohexane to cyclohexane/ethyl acetate1:1 as eluent affording the title compound (3 mg,) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.73 (2H, s), 7.07 (1H, t), 6.78 (1H, dd)6.52 (1H, dd) 5.69 (1H, br.s) 4.25-4.33 (2H, m) 1.95-2.05 (1H, m)1.74-1.86 (3H, m) 1.54-1.62 (5H, s), 1.00 (3H, t) 0.60-0.67 (2H, m).

Examples 36, Example 37 and Example 385,5-dimethyl-3-{6-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Racemate Mixture, Enantiomer 1, Enantiomer 2)

In a 50 mL round-bottomed flask the2-amino-2-methyl-N-[6-(4-methylchroman-5-yl)oxy-3-pyridyl]propanamide(Intermediate 215, 22.9 mg, 0.064 mmol) was dissolved in Dichloromethane(3 mL) to give a colourless solution. TEA (0.051 mL, 0.369 mmol) wasadded and the obtained mixture was cooled at 0° C. Triphosgene (21.87mg, 0.074 mmol) was dissolved in 1 ml of DCM and obtained solution wasadded to the reaction mixture. The reaction mixture was stirred at 0° C.for 15 minutes. Additional TEA (0.051 mL, 0.369 mmol) and triphosgene(21.87 mg, 0.074 mmol) dissolved in 1 ml of DCM were added to thereaction mixture at 0° C. and the reaction mixture was stirred for 15minutes. The reaction mixture was quenched with 5 mL of saturated sodiumbicarbonate solution and diluted with 10 mL of DCM. Phases wereseparated through a phase separator cartridge. The organic phase wasevaporated in vacuo and the residue was purified by flash chromatography(Biotage system) on silica gel using a column SNAP 10 g andcyclohexane/ethyl acetate from 2:1 to 1:2 as eluents affording the titlecompound Example 36 (19.1 mg) as a colorless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.30 (1H, d), 7.76 (1H, dd), 7.12 (1H, t),6.97 (1H, d), 6.72 (1H, dd), 6.61 (1H, dd), 5.74 (1H, br. s.), 4.23-4.32(1H, m), 4.15-4.22 (1H, m), 2.97-3.11 (1H, m), 2.07-2.16 (1H, m),1.62-1.70 (1H, m), 1.57 (6H, s), 1.28 (3H, d).

Both enantiomers of Example 36 were separated by preparative chiralchromatography.

Chiral preparative HPLC conditions were the following ones: Column:Chiralcel OD-H (25×2 cm), 5u; Mobile phase: n-Hexane/2-propanol 85:15%v/v; Flow rate: 18 ml/min; UV: 220 nm; Sample preparation: 18 mgdissolved in 1.2 ml of ethanol. Sample concentration: 15 mg/ml;Injection volume: 600 μL.

This preparative chiral chromatography afforded:

Example 37

7.1 mg of a white solid (enantiomer 1); Rt (Chiral preparativeHPLC)=21.018 minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.30 (1H, d), 7.76 (1H, dd), 7.12 (1H, t),6.97 (1H, d), 6.72 (1H, dd), 6.61 (1H, dd), 5.74 (1H, br. s.), 4.23-4.32(1H, m), 4.15-4.22 (1H, m), 2.97-3.11 (1H, m), 2.07-2.16 (1H, m),1.62-1.70 (1H, m), 1.57 (6H, s), 1.28 (3H, d).

Example 38

6.9 mg of a white solid (enantiomer 2); Rt (Chiral preparativeHPLC)=25.752 minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.30 (1H, d), 7.76 (1H, dd), 7.12 (1H, t),6.97 (1H, d), 6.72 (1H, dd), 6.61 (1H, dd), 5.74 (1H, br. s.), 4.23-4.32(1H, m), 4.15-4.22 (1H, m), 2.97-3.11 (1H, m), 2.07-2.16 (1H, m),1.62-1.70 (1H, m), 1.57 (6H, s), 1.28 (3H, d).

Examples 39, Example 40 and Example 41(5R)-5-ethyl-5-methyl-3-{6-[(3-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione(Diastereoisomeric Mixture, Diastereoisomer 1, Diastereoisomer 2)

In a 25 mL round-bottomed flask(2R)-2-amino-2-methyl-N-[6-(3-methylchroman-5-yl)oxy-3-pyridyl]butanamide(Intermediate 147, 33 mg, 0.088 mmol) was dissolved in Dichloromethane(5 mL) to give a colourless solution. The reaction mixture was cooled at0° C. TEA (0.061 mL, 0.441 mmol) and triphosgene (26.2 mg, 0.088 mmol)was added and the reaction mixture was stirred at 0° C. for 15 minutes.The reaction mixture was quenched with an aqueous saturated solution ofsodium bicarbonate (5 mL) and diluted with 10 mL of dichloromethane.Phases were separated through a phase separator cartridge. The organiclayer was evaporated in vacuo and the residue was purified by flashchromatography (Biotage system) on silica gel using a column SNAP 10 gand cyclohexane/ethyl acetate from 3:1 to 1:2 as eluents affording thetitle compound Example 39 (26.8 mg) as a yellow pale oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.26 (1H, d), 7.73 (1H, dd), 7.13 (1H, t),6.95 (1H, dd), 6.74 (1H, dd), 6.64 (1H, dd), 5.95 (1H, br. s.),4.15-4.21 (1H, m), 3.57-3.75 (1H, m), 2.70-2.84 (1H, m), 2.17 (2H, d),1.91-2.04 (1H, m), 1.70-1.82 (1H, m), 1.54 (3H, s), 1.01 (3H, d), 0.97(3H, t).

Both diastereoisomers of Example 39 were separated by Semipreparativechiral SFC.

Semipreparative chiral SFC conditions were the following ones: Column:Chiralpack AD-H (25×3 cm), 5 um; Modifier (Methanol+0.1% isopropylamine)20%; Flow rate 50 ml/min; Pressure 120 bar; Temperature 38° C.; UVdetection 220 nm; Loop 750 μL; Injection 13 mg (in Methanol).

This preparative chiral chromatography afforded:

Example 40

5.9 mg (diastereoisomer 1); Rt (Semipreparative chiral SFC)=13.48minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.26 (1H, d), 7.73 (1H, dd), 7.13 (1H, t),6.95 (1H, dd), 6.74 (1H, dd), 6.64 (1H, dd), 5.95 (1H, br. s.),4.15-4.21 (1H, m), 3.57-3.75 (1H, m), 2.70-2.84 (1H, m), 2.17 (2H, d),1.91-2.04 (1H, m), 1.70-1.82 (1H, m), 1.54 (3H, s), 1.01 (3H, d), 0.97(3H, t).

Example 41

6.6 mg (diastereoisomer 1); Rt (Semipreparative chiral SFC)=15.23minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.26 (1H, d), 7.73 (1H, dd), 7.13 (1H, t),6.95 (1H, dd), 6.74 (1H, dd), 6.64 (1H, dd), 5.95 (1H, br. s.),4.15-4.21 (1H, m), 3.57-3.75 (1H, m), 2.70-2.84 (1H, m), 2.17 (2H, d),1.91-2.04 (1H, m), 1.70-1.82 (1H, m), 1.54 (3H, s), 1.01 (3H, d), 0.97(3H, t).

Example 42 Example 43 and Example 44(5R)-5-ethyl-5-methyl-3-[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridinyl]-2,4-imidazolidinedione(Diastereoisomeric Mixture, Diastereoisomer 1, Diastereoisomer 2)

In a 25 mL round-bottomed flask(2R)—N-[6-(1,1a,2,7b-tetrahydrocyclopropa[c]chromen-7-yloxy)-3-pyridyl]-2-amino-2-methyl-butanamide(Intermediate 228, 33 mg, 0.088 mmol) was dissolved in Dichloromethane(5 mL) to give a colourless solution. The reaction mixture was cooled at0° C. TEA (0.091 mL, 0.66 mmol) and triphosgene (39 mg, 0.131 mmol) wereadded and the reaction mixture was stirred at 0° C. for 15 minutes. Thereaction mixture was quenched with an aqueous saturated solution ofsodium bicarbonate (5 mL) and diluted with 10 mL of dichloromethane.Phases were separated through a phase separator cartridge. The organiclayer was evaporated in vacuo and the residue was purified by flashchromatography (Biotage system) on silica gel using a column SNAP 10 gand cyclohexane/ethyl acetate from 3:1 to 1:2 as eluents affording thetitle compound Example 42 (42.6 mg) as a yellow pale oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.29 (1H, d), 7.73 (1H, dd), 7.07 (1H, t),6.97 (1H, d), 6.72 (2H, d), 5.77 (1H, br. s.), 4.31 (1H, dd), 3.95 (1H,dd), 1.92-2.11 (2H, m), 1.72-1.83 (1H, m), 1.64-1.73 (1H, m), 1.54 (3H,s), 0.97 (3H, t), 0.93-1.06 (2H, m).

Both diastereoisomers of Example 42 were separated by preparative chiralchromatography.

Preparative HPLC chiral chromatography conditions were the followingones: Column: Chiralpack AD-H (25×3 cm), 5 um; Mobile phase:n-Hexane/2-Propanol 90/10% v/v; Flow rate: 40 ml/min; UV: 220 nm; Samplepreparation: 41 mg dissolved in 4 ml of ethanol. Sample concentration:10.3 mg/ml; Injection volume: 2000 μL.

This preparative chiral chromatography afforded:

Example 43

14.1 mg as white solid (diastereoisomer 1); Rt (Chiral preparativeHPLC)=17.95 minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.29 (1H, d), 7.74 (1H, dd), 7.07 (1H, t),6.97 (1H, d), 6.72 (2H, d), 5.51 (1H, br. s.), 4.32 (1H, dd), 3.96 (1H,dd), 2.01-2.09 (1H, m), 1.92-2.02 (1H, m), 1.72-1.82 (1H, m), 1.64-1.73(1H, m), 1.55 (3H, s), 0.99-1.05 (2H, m), 0.95-1.01 (3H, m).

Example 44

15 mg as white solid (diastereoisomer 2); Rt (Chiral preparativeHPLC)=21.99 minutes

¹H NMR (400 MHz, CDCl₃) δ ppm 8.29 (1H, d), 7.74 (1H, dd), 7.07 (1H, t),6.97 (1H, d), 6.72 (2H, d), 5.51 (1H, br. s.), 4.32 (1H, dd), 3.96 (1H,dd), 2.01-2.09 (1H, m), 1.92-2.02 (1H, m), 1.72-1.82 (1H, m), 1.64-1.73(1H, m), 1.55 (3H, s), 0.99-1.05 (2H, m), 0.95-1.01 (3H, m).

Example 45, Example 46 and Example 473-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-5,5-dimethyl-2,4-imidazolidinedione(Racemate Mixture, Enantiomer 1, Enantiomer 2)

In a 50 mL round-bottomed flaskN1-{6-[(3-ethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-2-methylalaninamide(Intermediate 149, 17.2 mg, 0.050 mmol) was dissolved in Dichloromethane(3 mL) to give a pale yellow solution that was cooled at 0° C. TEA(0.035 mL, 0.252 mmol) was added followed by a dropwise addition of asolution of triphosgene (6.43 mg, 0.023 mmol) in dichloromethane (0.5ml) and the reaction mixture was stirred at 0° C. for 20 minutes. Thereaction mixture was evaporated in vacuo and the residue was purified byflash chromatography (Biotage system) on silica gel using a column SNAP10 g and cyclohexane/ethyl acetate from 2:1 to 1:2 as eluents affordingthe title compound Example 45 (15.3 mg) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ ppm 8.29 (1H, d), 7.80 (1H, dd), 7.33 (1H, t),7.09 (1H, d), 6.97-7.04 (2H, m), 5.70 (1H, br. s.), 5.23-5.30 (1H, m),5.19 (1H, dd), 5.12 (1H, d), 1.85-1.98 (1H, m), 1.67-1.80 (1H, m), 1.57(6H, s), 0.86-0.92 (3H, m).

Both enantiomers of Example 45 were separated by preparative chiralchromatography.

Chiral preparative HPLC conditions were the following ones: Column:Chiralpack AD-H (25×2 cm), 5u; Mobile phase: n-Hexane/2-propanol 85:15%v/v; Flow rate: 18 ml/min; UV: 220 nm; Sample preparation: 15 mgdissolved in 1.0 ml of ethanol (sample needs some drops of methanol fora complete solubilisation). Sample concentration: 15 mg/ml; Injectionvolume: 1000 μL.

This preparative chiral chromatography afforded:

Example 46

5.6 mg (enantiomer 1); Rt (Chiral preparative HPLC)=9.877 minutes

¹H NMR (400 MHz, MeOD) δ ppm 8.20 (1H, d), 7.92 (1H, dd), 7.38 (1H, t),7.18 (1H, d), 7.10 (1H, d), 7.02 (1H, d), 5.14-5.24 (2H, m), 5.09 (1H,d), 1.81-1.96 (1H, m), 1.65-1.79 (1H, m), 1.52 (6H, s), 0.86 (3H, s).

Example 47

5.5 mg (enantiomer 2); Rt (Chiral preparative HPLC)=13.203 minutes

¹H NMR (400 MHz, MeOD) δ ppm 8.20 (1H, d), 7.92 (1H, dd), 7.38 (1H, t),7.18 (1H, d), 7.10 (1H, d), 7.02 (1H, d), 5.14-5.24 (2H, m), 5.09 (1H,d), 1.81-1.96 (1H, m), 1.65-1.79 (1H, m), 1.52 (6H, s), 0.86 (3H, s).

Example 48, Example 49 and Example 50(5R)-5-ethyl-5-methyl-3-[2-(4-methylchroman-5-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione(Diastereoisomeric Mixture, Diastereoisomer 1, Diastereoisomer 2)

To a solution ofN1-{2-[(4-methyl-3,4-dihydro-2H-chromen-5-yl)oxy]-5-pyrimidinyl}-Disovalinamide(Intermediate 219, 78 mg, 0.219 mmol) in dry Dichloromethane (4 mL) TEA(0.076 mL, 0.547 mmol) was added. The mixture was cooled to 0° C. then asolution of triphosgene (29.2 mg, 0.098 mmol) in dry Dichloromethane(1.0 mL) was added dropwise. The reaction mixture was stirred for 10minutes at 0° C. then it was maintained in the ice-bath and quenchedwith water (10 mL). The organic layer was collected and the aqueousphase was extracted with DCM (2×5 mL). The combined organic layers weredried (Na₂SO₄), filtered and evaporated. The residue was purified byflash chromatography on silica gel using a column SNAP 25 g andcyclohexane/ethyl acetate from 8:2 to 1:1 as eluents affording the titlecompound (66 mg) as a white solid.

UPLC_IPQC: 0.99 min, 383 [M+H]+.

Both diastereoisomers of Example 48 were separated by Semipreparativechiral SFC.

Semipreparative chiral SFC conditions were the following ones: Column:Chiralpack AD-H (25×0.46 cm), 5 um; Modifier (Ethanol+0.1%isopropylamine) 20%; Flow rate 2.5 ml/min; Pressure 120 bar; Temperature38° C.; UV detection 210-340 nm.

This preparative chiral chromatography afforded:

Example 49

24 mg as white solid (diastereoisomer 1); Rt (Chiral preparativeSFC)=7.583 min

¹H NMR (400 MHz, CDCl₃) δ ppm 8.72 (2H, s), 7.17 (1H, t), 6.79 (1H, d),6.68 (1H, d), 5.36 (1H, br.s), 4.25-4.35 (1H, m), 4.14-4.25 (1H, m),2.98-3.09 (1H, m), 2.06-2.20 (1H, m), 1.95-2.06 (1H, m), 1.74-1.87 (1H,m), 1.70 (1H, d), 1.59 (3H, s), 1.31 (3H, s), 1.01 (3H, t).

Example 50

25 mg as white solid (diastereoisomer 2); Rt (Chiral preparativeSFC)=10.156 min

¹H NMR (400 MHz, CDCl₃) δ ppm 8.72 (2H, s), 7.17 (1H, t), 6.78 (1H, d),6.68 (1H, d), 5.38 (1H, br.s), 4.25-4.36 (1H, m), 4.12-4.25 (1H, m),2.94-3.12 (1H, m), 2.05-2.20 (1H, m), 1.95-2.05 (1H, m), 1.75-1.87 (1H,m), 1.63-1.74 (1H, m), 1.58 (3H, s,) 1.31 (3H, s), 1.01 (3H, t).

Example 51(5R)-5-ethyl-5-methyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione

To a solution of 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 156, 18 mg, 0.1 mmol) in dry DMF (1 ml) potassiumcarbonate (27.6 mg, 0.2 mmol) and then(5R)-3-(2-chloropyrimidin-5-yl)-5-ethyl-5-methyl-imidazolidine-2,4-dione(Intermediate 165, 20 mg, 0.08 mmol) were added and the reaction mixturewas stirred for 2 hours at 80° C. After cooling the reaction mixture wasquenched with water (1 ml), diluted with brine (5 ml) and extracted withethyl acetate (2×10 ml). The organic layer was dried over sodiumsulfate, filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 7:3 to cyclohexane/ethyl acetate 3:7 aseluents affording the title compound (21 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm: 8.69-8.74 (3H, m), 6.94 (1H, d), 6.52(1H, d), 4.44 (2H, s), 2.15 (3H, s), 1.73-1.83 (1H, m), 1.63-1.73 (1H,m), 1.40 (3H, s), 1.02-1.06 (2H, m), 0.85-0.92 (5H, m). LC/MS:QC_(—)3_MIN: Rt=2.007 min; 395 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 152) with the appropriate phenol. Final products werepurified by flash-chromatography (Silica cartridge; Cyclohexane/EtOAc orother appropriate solvent system).

Ex. Structure Name Phenol ¹H-NMR LCMS 52

(5R)-3-[2-(3,3- dimethyl- isochroman-5- yl)oxypyrimidin-5-yl]-5-ethyl-5- methyl- imidazolidine- 2,4-dione 3,3- dimethyliso-chroman-5- ol (Inter- mediate 170) ¹H-NMR (400 MHz, DMSO-d₆) δ ppm:8.70-8.75 (3H, m), 7.27 (1H, t), 7.06 (2H, dd), 4.75 (2H, s), 2.42 (2H,s), 1.63- 1.75 (2H, m), 1.41 (3H, s), 1.18 (6H, s), 0.88 (3H, t). LC/MS:QC_3_MIN: Rt = 1.894 min; 397 [M + H]+. 53

(5R)-5-ethyl-5- methyl-3-[2-(7- methylspiro[1H- isobenzofuran- 3,1′-cyclobutane]-4- yl)oxypyrimidin- 5- yl]imidazolidine- 2,4-dione 7-methyl- spiro[1H- isobenzo- furan-3,1′- cyclo- butane]-4- ol (Inter-mediate 176) ¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.77-8.73 (3H, m), 7.16(1H, d), 7.07 (1H, d), 4.98 (2H, s), 2.49- 2.28 (4H, m), 2.21 (3H, s),1.89-1.62 (4H, m), 1.41 (3H, s), 0.93-0.86 (3H, m). LC/MS: QC_3_MIN: Rt= 1.997 min 409 [M + H]+. 54

(5R)-5-ethyl-5- methyl-3-{2- [(3,3,7-trimethyl- 2,3-dihydro-1-benzofuran-4- yl)oxy]-5- pyrimidinyl}-2,4- imidazolidine- dione 3,3,7-trimethyl- 2,3- dihydro-1- benzo- furan- 4-ol (Inter- mediate 184) ¹HNMR (400 MHz, DMSO-d₆) δ ppm: 8.70 (1H, br.s), 8.69 (2H, s), 6.98 (1H,d), 6.55 (1H, d), 4.19 (2H, s), 2.12 (3H, s), 1.90-1.50 (1H, m), 1.38(3H, s), 1.21 (6H, s), 0.86 (3H, t). UPLC: 1.06 min, 397 [M + H ]+ 55

(5R)-3-{2-[(2,2- difluoro-7- methyl-1,3- benzodioxol-4- yl)oxy]-5-pyrimidinyl}-5- ethyl-5-methyl- 2,4- imidazolidine- dione 2,2-difluoro-7- methyl-1,3- benzo- dioxol-4- ol (Inter- mediate 197) ¹H-NMR(400 MHz, DMSO-d₆): δ ppm 8.78 (2H, s), 8.74 (1H, br.s), 7.18-7.13 (2H,m), 2.33 (3H, s), 1.84- 1.75 (1H, m), 1.71-1.62 (1H, m), 1.40 (3H, s),0.88 (3H, t. UPLC: 1.11 min, 407 [M + H]+, 56

(5R)-3-{2-[(2,2- difluoro-1,3- benzodioxol-4- yl)oxy]-5- pyrimidinyl}-5-ethyl-5-methyl- 2,4- imidazolidine- dione 2,2- difluoro- 1,3- benzo-dioxol- 4-ol (Inter- mediate 198) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.80(2H, s), 8.75 (1H, s), 7.39 (1H, d), 7.31 (1H, t), 7.27 (1H, d), 1.84-1.75 (1H, m), 1.72-1.62 (1H, m), 0.88 (3H, t). UPLC: 1.04 min, 393 [M +H]+ 57

(5R)-5-ethyl-5- methyl-3-{2- [(2,4,4-trimethyl- 4H-3,1- benzoxazin-5-yl)oxy]-5- pyrimidinyl]-2,4- imidazolidine- dione 2,4,4- trimethyl-4H-3,1- benzoxazin- 5-ol (Inter- mediate 202) ¹H NMR (400 MHz, DMSO-d₆):δ ppm 8.73 (2H, s), 8.71 (1H, s), 7.30- 7.25 (1H, m), 6.96 (2H, ddd),2.01 (3H, s), 1.82- 1.58 (2H, m), 1.51 (6H, s), 1.38 (3H, s), 0.85 (3H,t).

Example 585,5-dimethyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione

To a solution of 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 156, 18 mg, 0.1 mmol) in dry DMF (1 ml) potassiumcarbonate (27.6 mg, 0.2 mmol) and then3-(2-chloropyrimidin-5-yl)-5,5-dimethyl-imidazolidine-2,4-dione(Intermediate 166, 20 mg, 0.083 mmol) were added and the reactionmixture was stirred for 2 hours at 80° C. After cooling the reactionmixture was quenched with water (1 ml), diluted with brine (5 ml) andextracted with ethyl acetate (2×10 ml). The organic layer was dried oversodium sulfate, filtered and evaporated and the residue was purified byflash chromatography (Biotage system) on silica gel using a 10 g SNAPcolumn and cyclohexane/ethyl acetate 7:3 to cyclohexane/ethyl acetate3:7 as eluents affording the title compound (18 mg) as a light beigesolid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.74 (1H, s), 8.70 (2H, s), 6.94 (1H,d), 6.52 (1H, d), 4.44 (2H, s), 2.14 (3H, s), 1.42 (6H, s), 1.01-1.06(2H, m), 0.87-0.92 (2H, m). LC/MS: QC_(—)3_MIN: Rt=1.946 min; 380[M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol(Intermediate 152) with the appropriate phenol. Final products werepurified by flash-chromatography (Silica cartridge; Cyclohexane/EtOAc orother appropriate solvent system).

Ex. Structure Name Phenol 1H-NMR LCMS 59

3-[2-(3,3- dimethyl- isochroman- 5-yl) oxypyrimidin- 5-yl]-5,5-dimethyl- imidazolidine- 2,4-dione 3,3- dimethyl- isochroman- 5-ol(Inter- mediate 170) ¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.75 (1H, s), 8.71(1H, s), 7.28 (1H, t), 7.06 (1H, dd), 4.75 (2H, s), 2.41 (2H, s), 1.44(6H, s), 1.18 (6H, s). LC/MS: QC_3_MIN: Rt = 1.822 min; 383 [M + H]+. 60

5,5-dimethyl-3-[2- (7-methylspiro[1H- isobenzofuran- 3,1′-cyclobutane]-4-yl)oxypyrimidin- 5-yl]imidazolidine- 2,4-dione 7- methyl- spiro[1H-isobenzo- furan-3,1′- cyclo- butane]-4- ol (Inter- mediate 176) ¹H-NMR(400 MHz, DMSO-d₆) δ ppm: 8.79- 8.73 (3H, m), 7.17 (1H, d), 7.07 (1H,d), 4.98 (2H, s), 2.49-2.28 (4H, m), 2.21 (3H, s), 1.89-1.67 (2H, m),1.43 (6H, s). LC/MS: QC_3_MIN: Rt = 1.936 min 395 [M + H]+.

Example 61(5R)-5-ethyl-3-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]imidazolidine-2,4-dione

To a solution of(2R)-2-amino-N-[2-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyrimidin-5-yl]butanamide(Intermediate 164, 24 mg, 0.068 mmol) in dry DCM (3 ml) TEA (0.028 ml,0.2 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (15 mg, 0.05 mmol) in dry DCM (1.5 ml) wasslowly added and the reaction mixture was stirred for 15 minutes at thesame temperature. The reaction was quenched with water (10 ml) and twophases were separated. The organic layer was dried (Na₂SO₄), filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 10 g SNAP column andcyclohexane/ethyl acetate 75:25 to cyclohexane/ethyl acetate 25:75 aseluents affording the title compound (11 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm: 8.75 (1H, s), 8.68 (2H, s), 6.94 (1H,d), 6.52 (1H, d), 4.44 (2H, s), 4.20-4.25 (1H, m), 2.15 (3H, s),1.77-1.88 (1H, m), 1.66-1.76 (1H, m), 1.02-1.06 (2H, m), 0.96 (3H, t),0.87-0.92 (2H, m). LC/MS: QC_(—)3_MIN: Rt=1.955 min; 381 [M+H]+.

Example 62(5R)-5-ethyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione

To a solution of(2R)-2-amino-N-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]butanamide(Intermediate 160, 40 mg, 0.11 mmol) in dry DCM (5 ml) TEA (0.042 ml,0.3 mmol) was added and the reaction mixture was cooled to 0° C. Asolution of triphosgene (23.7 mg, 0.08 mmol) in dry DCM (3 ml) wasslowly added and the reaction mixture was stirred for 15 minutes at thesame temperature. The reaction was quenched with water (10 ml) and twophases were separated. The organic layer was dried (Na₂SO₄), filteredand evaporated and the residue was purified by flash chromatography(Biotage system) on silica gel using a 10 g SNAP column andcyclohexane/ethyl acetate 75:25 to cyclohexane/ethyl acetate 25:75 aseluents affording the title compound (22 mg) as a white solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.63 (1H, s), 8.13 (1H, d), 7.84 (1H,dd), 7.07 (1H, d), 6.94 (1H, d), 6.44 (1H, d), 4.46 (2H, s), 4.19-4.24(1H, m), 2.15 (3H, s), 1.77-1.88 (1H, m), 1.65-1.75 (1H, m), 1.10-1.14(2H, m), 0.96 (3H, t), 0.87-0.92 (2H, m). LC/MS: QC_(—)3_MIN: Rt=2.025min; 380 [M+H]+.

Example 63(5R)-5-ethyl-3-{6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}-2,4-imidazolidinedione

(2R)-2-amino-N-{6-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-3-pyridinyl}butanamide(Intermediate 188, 300 mg, 0.84 mmol) was dissolved in ethyl acetate (6mL). Triethylamine (0.47 ml, 3.36 mmol) was added and the reactionmixture was cooled to 0° C. A solution of triphosgene (100 mg, 0.34mmol) in ethyl acetate (6 mL) was slowly added. At the end of additionthe mixture was treated with an aqueous saturated solution of NaHCO₃ andtwo phases were separated. The organic layer was washed with brine,dried over Na₂SO₄ and evaporated to dryness to obtain a waxy solid. Theresidue was purified by flash chromatography on silica gel usingcyclohexane/ethyl acetate 70:30 to cyclohexane/ethyl acetate 50:50 aseluents affording the title compound (166 mg) as a white foam.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.61 (1H, br.s), 8.12 (1H, d), 7.82(1H, dd), 7.10 (1H, d), 6.98 (1H, d), 6.47 (1H, d), 4.21 (2H, s), 4.18(1H, br.s), 2.13 (3H, s), 1.86-176 (1H, m), 1.75-1.64 (1H, m), 1.25 (6H,s), 0.95 (3H, t). ¹³C-NMR (200 MHz, DMSO-d₆): δ ppm 173.2, 162.5, 158.6,155.4, 148.2, 145.2, 138.5, 130.0, 126.1, 124.3, 115.7, 114.4, 110.6,83.6, 57.5, 42.2, 26.0, 24.4, 14.4, 8.8.

Example 64(5R)-5-ethyl-3-{2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}-2,4-imidazolidinedione

1,1-dimethylethyl{(1R)-1-[({2-[(3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-yl)oxy]-5-pyrimidinyl}amino)carbonyl]propyl}carbamate(Intermediate 191, 213 mg, 0.47 mmol) was dissolved in HCl 5-6 N inisopropanol (1 mL) and the resulting solution was heated to 35° C. for30 minutes. The reaction mixture was then concentrated under vacuum, theresidue diluted with ethyl acetate (50 mL) and an aqueous 5% solution ofK₂CO₃ (30 mL). Two phases were separated and the organic layer waswashed with an aqueous saturated solution of ammonium chloride (30 mL),dried over Na₂SO₄ and concentrated under vacuum. The resulting crude wasdissolved in ethyl acetate (10 mL) and triethylamine was added (0.23 mL,1.64 mmol). The reaction mixture was cooled to 0-5° C. and a solution oftriphosgene (55 mg, 0.185 mmol) in ethyl acetate (5 mL) was added dropwise in 10 minutes. The reaction was quenched with water (50 mL) andextracted with ethyl acetate (50 mL). The organic layer was washed withbrine dried over Na₂SO₄ and concentrated under vacuum. The residue waspurified purified by flash chromatography on silica gel usingcyclohexane/ethyl acetate 50:50 as eluent affording the title compound(161 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.72 (1H, s), 8.66 (2H, s), 7.03-6.93(1H, m), 6.55 (1H, d), 4.18 (2H, s), 2.12 (3H, s), 1.87-1.61 (2H, m),1.2 (6H, s), 1.15 (1H, t), 0.94 (3H, t). MS_(—)2 (ESI): 383 [M+H].

Example 65(5R)-5-ethyl-5-methyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione

To a solution of triphosgene (30 mg, 0.1 mmol) in dry DCM (1 ml) at 0°C., under nitrogen atmosphere, DIPEA (0.175 ml, 1.0 mmol) was addedfollowed by the addition (slowly added) of a solution of6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyridin-3-amine(Intermediate 158, 27 mg, 0.1 mmol) in dry DCM (2 ml) and the reactionmixture was stirred for 15 minutes at the same temperature. After that asolution of Methyl (R)-2-amino-2-methyl-butyrate hydrochloride (33 mg,0.2 mmol) in dry DCM (2 ml) was added and the reaction mixture wasstirred for 30 minutes at 0° C. The reaction was quenched with a 1Maqueous solution of HCl (5 ml), diluted with DCM (10 ml) and two phaseswere separated. The organic layer was washed with brine (10 ml), dried(Na₂SO₄), filtered and evaporated affording the urea intermediate asyellow foam.

The urea was dissolved in MeOH (5 ml), NaOMe (10 mg) was added and thereaction mixture was stirred for 15 minutes at room temperature. Thereaction was quenched with an aqueous saturated solution of ammoniumchloride (20 ml) and diluted with ethyl acetate (40 ml). Two phases wereseparated and the organic layer was dried (Na₂SO₄), filtered andevaporated and the residue was purified by flash chromatography (Biotagesystem) on silica gel using a 10 g SNAP column and cyclohexane/ethylacetate 75:25 to cyclohexane/ethyl acetate 25:75 as eluents affordingthe title compound (29 mg) as a white solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.60 (1H, s), 8.15 (1H, d), 7.85 (1H,dd), 7.06 (1H, d), 6.94 (1H, d), 6.44 (1H, d), 4.46 (2H, s), 2.15 (3H,s), 1.73-1.83 (1H, m), 1.62-1.72 (1H, m), 1.40 (3H, s), 1.10-1.14 (2H,m), 0.84-0.92 (5H, m). LC/MS: QC_(—)3_MIN: Rt=2.076 min; 394 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyridin-3-amine(Intermediate 154) with the appropriate aniline. Final products werepurified by flash-chromatography (Silica cartridge; Cyclohexane/EtOAc orother appropriate solvent system).

Ex. Structure Name Aniline ¹H-NMR LCMS 66

(5R)-3-[6-(3,3- dimethyl- isochroman- 5-yl) oxy-3- pyridyl]-5- ethyl-5-methyl- imidazolidine- 2,4-dione 6-(3,3- dimethyl- isochroman- 5-yl)oxy-pyridin-3- amine (Inter- mediate 172) ¹H-NMR (400 MHz, DMSO-d₆) δ ppm:8.11 (1H, d), 7.86 (1H, dd), 7.25 (1H, t), 7.13 (1H, d), 7.00 (2H, dd),4.75 (2H, s), 2.44 (2H, s), 1.61- 1.84 (2H, m), 1.40 (3H, s), 1.18 (6H,s) 0.87 (3H, t). LC/MS: QC_3_MIN: Rt = 1.962 min; 396 [M + H]+. 67

(5R)-3-[6- [(3,3-diethyl- 1H- isobenzo- furan-4-yl) oxy]-3- pyridyl]-5-ethyl-5- methyl- imidazolidine- 2,4-dione 6-[(3,3- diethyl-1H- isobenzo-furan-4- yl)oxy] pyridin-3- amine (Inter- mediate 180) ¹H-NMR (400 MHz,DMSO-d₆) δ ppm: 8.61 (1H, s), 8.16 (1H, d), 7.89- 7.87 (1H, dd), 7.35(1H, t), 7.18 (1H, d), 7.14 (1H, d), 7.10 (1H, d), 5.08 (2H, s),1.85-1.61 (6H, m), 1.40 (3H, s), 0.88 (3H, t), 0.67 (6H, t). LC/MS:QC_3_MIN: Rt 2.067 min, 410 [M + H]+.

Example 68(5R)-5-ethyl-5-methyl-3-[6-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]-3-pyridyl]imidazolidine-2,4-dione

To a solution of(2R)-2-amino-2-methyl-N-[6-[(2,4,4-trimethyl-3,1-benzoxazin-5-yl)oxy]-3-pyridyl]butanamide(Intermediate 205, 143 mg, 0.37 mmol) and TEA (0.21 mL) in ethyl acetate(2.5 mL) at 0° C., triphosgene (44 mg, 0.15 mmol) dissolved in ethylacetate (2.5 mL) was added drop wise in 10 min. The reaction mixture wasstirred at 0° C. for 40 min. The reaction mixture was diluted with water(5 mL) and ethyl acetate (10 mL). Phases were separated and the aqueouswas back-extracted with ethyl acetate (2×10 mL). The combined organiclayers were washed with brine (15 mL), dried over Na₂SO₄, filtered andconcentrated and the residue was purified by flash chromatography onsilica gel using ethyl acetate as eluent affording the title compound(115 mg) as white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.58 (1H, s), 8.15 (1H, d), 7.87 (1H,dd), 7.24 (1H, t), 7.17 (1H, d), 6.89 (1H, d), 6.84 (1H, d), 2.01 (3H,s), 1.81-1.70 (1H, m), 1.69-1.59 (1H, m), 1.54 (6H, s), 1.37 (3H, s),0.84 (3H, t).

Example 69(5R)-3-{6-[(3,3-dimethyl-1,3-dihydro-2-benzofuran-4-yl)oxy]-3-pyridinyl}-5-ethyl-5-methyl-2,4-imidazolidinedione

3,3-Dimethyl-1,3-dihydro-2-benzofuran-4-ol (Intermediate 193, 68 mg, 0.4mmol) and(5R)-3-(6-chloropyridin-3-yl)-5-ethyl-5-methylimidazolidine-2,4-dione(Intermediate 194, 126 mg, 0.48 mmol) were dissolved in DMF (1.0 mL) andK₂CO₃ (143 mg, 1.03 mmol) was added. The resulting suspension was heatedto 130° C. under microwave irradiation for 40 minutes. The reactionmixture was diluted with water (25 mL) and ethyl acetate (25 mL), andthen two phases were separated. The organic layer was washed with waterand brine, dried over Na₂SO₄ and concentrated under vacuum. The residuewas purified by flash chromatography (Biotage system) on silica gelusing cyclohexane/ethyl acetate 50:50 as eluent affording the titlecompound (70 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.57 (1H, s), 8.12 (1H, d), 7.86 (1H,dd), 7.30 (1H, t), 7.22-7.03 (2H, m), 6.96 (1H, d), 4.99 (2H, s),1.80-1.57 (2H, m), 1.37 (6H, s), 1.36 (3H, s), 0.84 (3H, s). MS_(—)2(ESI): 382 [M+H]+.

Example 705,5-dimethyl-3-[6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxy-3-pyridyl]imidazolidine-2,4-dione

To a solution of triphosgene (30 mg, 0.1 mmol) in dry DCM (1 ml) at CPC,under nitrogen atmosphere, DIPEA (0.175 ml, 1.0 mmol) was added followedby the addition (slowly added) of a solution of6-(7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-yl)oxypyridin-3-amine(Intermediate 158, 27 mg, 0.1 mmol) in dry DCM (2 ml) and the reactionmixture was stirred for 15 minutes at the same temperature. After that asolution of Methyl 2-amino-2-methylpropanoate hydrochloride (30 mg, 0.2mmol) in dry DCM (2 ml) was added and the reaction mixture was stirredfor 30 minutes at 0° C. The reaction was quenched with a 1M aqueoussolution of HCl (5 ml), diluted with DCM (10 ml) and two phases wereseparated. The organic layer was washed with brine (10 ml), dried(Na₂SO₄), filtered and evaporated affording the urea intermediate asyellow foam.

The urea was dissolved in MeOH (5 ml), NaOMe (10 mg, 0.19 mmol) wasadded and the reaction mixture was stirred for 15 minutes at roomtemperature. The reaction was quenched with an aqueous saturatedsolution of ammonium chloride (20 ml) and diluted with ethyl acetate (40ml). Two phases were separated and the organic layer was dried (Na₂SO₄),filtered and evaporated and the residue was purified by flashchromatography (Biotage system) on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate 75:25 to cyclohexane/ethyl acetate 25:75as eluents affording the title compound (23 mg) as a white solid.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.62 (1H, s), 8.14 (1H, d), 7.86 (1H,dd), 7.05 (1H, d), 6.92 (1H, d), 6.43 (1H, d), 4.44 (2H, s), 2.14 (3H,s), 1.40 (6H, s), 1.08-1.13 (2H, m), 0.96 (3H, t), 0.85-0.90 (2H, m).LC/MS: QC_(—)3_MIN: Rt=2.016 min; 380 [M+H]+.

The following Reference Intermediates and Examples describe thepreparation of compounds for use in assays.

Reference Intermediate R1 4-methyl-3-(methyloxy)aniline

To a solution of 1-methyl-2-(methyloxy)-4-nitrobenzene (2.5 g, 14.96mmol) in methanol (50 mL) Ni-Raney (˜2 g) was added and the reactionmixture was stirred overnight at room temperature under H₂ atmosphere (1atm). The catalyst was filtered off and the residue was purified by SCXcartridge (50 g) to afford the title compound (1.86 g) as a colourlessoil.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 6.73 (1H, d), 6.19 (1H, d), 6.05 (1H,dd), 4.85 (2H, s), 3.68 (3H, s), 1.97 (3H, s); UPLC_B: 0.62 min, 138[M+H]+.

Reference Intermediate R2 4-methyl-3-(methyloxy)phenol

To a suspension of 4-methyl-3-(methyloxy)aniline (Reference IntermediateR1, 1.86 g) in water (100 mL)/H₂SO₄ (30 mL, 563 mmol) at 0° C. asolution of sodium nitrite (1.029 g, 14.91 mmol) in water (10 mL) wasslowly added and the reaction mixture was stirred for 30 minutes at 0°C. The reaction mixture was slowly added to a solution of H₂SO₄ 98% (20mL) in Water (80 mL) pre-heated at 90° C. and stirred at thistemperature for 1 h. After cooling the mixture was extracted with Et₂O(2×200 mL), the organic layer was dried on sodium sulphate, filtered andevaporated to afford the title compound (1.86 g) as a red/brown oil.

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 9.14 (1H, br.s), 6.87 (1H, d), 6.35(1H, d), 6.24 (1H, dd), 3.71 (3H, s), 2.01 (3H, s); UPLC_B: 0.63 min,137 [M−H]−.

Reference Intermediate R32-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine

To a solution of 4-methyl-3-(methyloxy)phenol (Reference IntermediateR2, 400 mg) in dry N,N-dimethylformamide (15 mL), potassium carbonate(1200 mg, 8.69 mmol) and then 2-chloro-5-nitropyridine (551 mg, 3.47mmol) were added and the reaction mixture was stirred for 2 hours at115° C. The reaction was quenched with water (10 mL), diluted with brine(20 mL) and extracted with ethyl acetate (3 times 30 mL). The organiclayer was washed with ice cold brine (2 times 30 mL), dried over sodiumsulphate, filtered and evaporated. The residue was purified by silicagel chromatography (Biotage system, 100 g SNAP column) with a gradientcyclohexane/ethyl acetate from 10/0 to 8/2. Evaporation afforded the totitle compound as a light yellow oil (570 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 2.16 (3H, s), 3.76 (3H, s), 6.68-6.73(1H, m), 6.83-6.86 (1H, m), 7.24-7.18 (2H, m), 8.64-8.58 (1H, m),9.08-9.02 (1H, m); UPLC_B: 0.93 min, 261 [M+H]+.

Reference Intermediate R46-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine

To a solution of 2-{[4-methyl-3-(methyloxy)phenyl]oxy}-5-nitropyridine(Reference Intermediate R3, 568 mg) in tetrahydrofuran (25 mL)/water(12.50 mL), iron (609 mg, 10.91 mmol) and then ammonium chloride (584mg, 10.91 mmol) were added and the reaction mixture was stirred for 8hours at room temperature. The catalyst was filtered off and thesolution was diluted with an aqueous saturated solution of Na₂CO₃ (5 mL)and extracted with ethyl acetate (2 times 40 mL). Combined organiclayers were dried over sodium sulphate, filtered and evaporated and theresidue was purified by silica gel chromatography (Biotage system with a50 g SNAP column) using a as eluent a gradient cyclohexane/ethyl acetatefrom 8/2 to 1/1. Evaporation afforded the title compound as light yellowoil (465 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 7.54 (1H, d), 7.06 (2H, ddd), 6.72 (1H,d), 6.59 (1H, d), 6.38 (1H, dd), 5.07 (2H, s), 3.73 (3H, s), 2.10 (3H,s); UPLC_B: 0.72 min, 231 [M+H]+.

Reference Intermediate R51,1-dimethylethyl((1R)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (106 mg,0.521 mmol) in dry N,N-dimethylformamide (2 mL) DIPEA (0.152 mL, 0.869mmol) and then TBTU (181 mg, 0.565 mmol) were added and the reactionmixture was stirred for 15 minutes at room temperature.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (ReferenceIntermediate R4, 100 mg) was then added and the reaction mixture wasstirred overnight at the same temperature. The reaction was quenchedwith water (1 mL), diluted with brine (1 mL) and extracted with ethylacetate (3 times 5 mL). The organic layer was dried over sodiumsulphate, filtered and evaporated and the residue was purified by silicagel chromatography (Biotage system, 10 g SNAP column) using as eluent agradient cyclohexane/ethyl acetate from 100/0 to 70/30 to afford thetitle compound as a white solid (180 mg).

¹H NMR (400 MHz, DMSO-d₆) δ ppm: 10.13 (1H, br. s), 8.31-8.37 (1H, m),8.02-8.10 (1H, m), 7.09-7.16 (1H, m), 7.01-7.08 (1H, m), 6.96 (1H, d),6.70 (1H, d), 6.51-6.58 (1H, m), 3.91-4.03 (1H, m), 3.75 (3H, s), 2.13(3H, s), 1.50-1.76 (2H, m), 1.39 (9H, s), 0.90 (3H, t); UPLC_B: 0.91min, 416 [M+H]+.

Reference Intermediate R6(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide

To a solution of1,1-dimethylethyl((1R)-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Reference Intermediate R5, 175 mg) in dry dichloromethane (DCM) (6 mL)TFA (2 mL, 26.0 mmol) was slowly added and the reaction mixture wasstirred for 1 h at room temperature. The solvent and the excess of TFAwere evaporated and the residue was purified by SCX cartridge (5 g) toafford the title compound as a colourless solid (122 mg).

¹H-NMR (400 MHz, DMSO-d₆) δ ppm: 8.36-8.42 (1H, m), 8.11 (1H, dd), 7.12(1H, d), 6.95 (1H, d), 6.67-6.73 (1H, m), 6.54 (1H, dd), 3.75 (3H, s),3.24 (1H, m), 2.13 (3H, s), 1.59-1.73 (1H, m), 1.42-1.56 (1H, m), 0.90(3H, t); UPLC_B: 0.74 min, 316 [M+H]+.

Reference Intermediate R71,1-dimethylethyl((1R)-1-methyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of N-{[(1,1-dimethylethyl)oxy]carbonyl}-D-isovaline (94mg, 0.434 mmol) in dry N,N-Dimethylformamide (1 mL) DIPEA (0.114 mL,0.651 mmol) and HATU (165 mg, 0.434 mmol) were added. The reaction wasstirred at room temperature for 15 minutes.6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinamine (ReferenceIntermediate R4, 50 mg) was then added. After 1 hour of stirring at roomtemperature the mixture was heated at 50° C. and stirred at thattemperature for 4 hours, it was then cooled down to room temperature andstirred overnight at that temperature. The mixture was quenched withbrine (2 mL) and extracted with ethyl acetate (3×2 mL). Combined organiclayers were dried over sodium sulphate, filtered and evaporated. Theresidue was purified by flash chromatography on silica gel using a 10 gSNAP column and cyclohexane/ethyl acetate as eluents from 100/0 to 60/40(Biotage system) to afford the title compound as a white solid (65 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.60 (1H, br s), 9.10 (1H, br s), 8.31(1H, br. s.), 8.03 (1H, br s), 7.12 (1H, d), 6.93 (1H, d), 6.69 (1H, d),6.53 (1H, dd), 3.74 (3H, s), 2.11 (3H, s), 1.72-1.86 (1H, m), 1.60-1.72(1H, m), 1.41 (9H, s), 1.33 (3H, s), 0.78 (3H, t); UPLC: 0.87 min, 430[M+H]+

Reference Intermediate R8N¹-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-isovalinamide

To a solution of1,1-dimethylethyl((1R)-1-methyl-1-{[(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Reference Intermediate R7, 65 mg) in dry dichloromethane (3 mL) cooledto 0° C., TFA (0.700 mL, 9.08 mmol) was added dropwise. The reaction wasstirred at that temperature for 2 hours. The reaction was quenched witha saturated aqueous solution of NaHCO₃ (20 mL) added at 0° C., andextracted with dichloromethane (3×7 mL). The combined organic layerswere dried over sodium sulphate, filtered and evaporated to afford thetitle compound as a white solid (44 mg).

Reference Intermediate R9 3-(1,1-dimethylethyl)-4-hydroxybenzaldehyde

2-(1,1-dimethylethyl)phenol (10 g, 66.67 mmol) was dissolved in 40 mL ofMeOH and NaOH (40 g, 1 mol) dissolved in 40 mL of water was addeddropwise. Then 40 mL of CHCl₃ was added (during the course of 1 h) at60° C. The reaction mixture was stirred at that temperature for 3 h.After cooling down to r.t., the mixture was cooled to 0° C. and 4M HClwas added until the solution reached pH 5-6. The mixture was extractedwith DCM (three times) and the collected organic were dried over Na₂SO₄,filtered and evaporated. The crude was charged on a silica gel columnand eluted with Cyclohexane/EtOAc (from 100:0 to 80:20Cyclohexane/EtOAc, then plateau at 80:20) affording 766 mg of the of thetitle compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.62 (1H, s), 9.79 (1H, s), 7.73 (1H,br. s), 7.67-7.57 (1H, m), 7.01-6.90 (1H, m), 1.38 (9H, s); UPLC_ipqc:0.97 min, 177 [M−H]⁻.

Reference Intermediate R10 3-(1,1-dimethylethyl)-4-hydroxybenzonitrile

3-(1,1-dimethylethyl)-4-hydroxybenzaldehyde (Reference Intermediate R9,550 mg) and hydroxylamine hydrochloride (322 mg, 4.63 mmol) were stirredin 8.0 mL of acetic acid at reflux for 1 h. After cooling down to 0° C.,the mixture was poured into Et₂O and washed once with water and oncewith NaOH (5% aqueous solution). The collected aqueous phases wereextracted with Et₂O (two times) and the combined organic phases weredried over Na₂SO₄, filtered, evaporated and triturated with pentaneaffording 540 mg of the title compound.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.92 (1H, br. s), 7.53-7.45 (2H, m),6.92 (1H, d), 1.34 (9H, s); UPLC_ipqc: 1.03 min, 174 [M−H]⁻.

Reference Intermediate R11 4-hydroxy-2-iodobenzonitrile

To a solution of 2-fluoro-4-iodobenzonitrile (5.0 g, 20.24 mmol) in dryacetonitrile (100 mL) potassium trimethylsilanolate (1.18 g) was addedand the reaction mixture was stirred overnight at 50° C. The solvent wasremoved under reduced pressure and the residue was dissolved in ethylacetate (100 mL) and an aqueous pH 3 buffer solution was added up topH˜5. Two phases were separated and the organic layer was dried(Na₂SO₄), filtered and evaporated to afford the title compound (4.90 g)as brown solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 10.92 (1H, s), 7.65 (1H, d), 7.39 (1H,d), 6.93 (1H, dd); UPLC_ipqc: 0.81 min, 244 [M−H]−.

Reference Intermediate R124-hydroxy-2-[(trifluoromethyl)oxy]benzonitrile

Two reactions were carried out in parallel (A and B) and then the tworeaction mixtures were combined to run work-up and purification.

Reaction A: To a solution of 4-Methoxy-2-(trifluoromethoxy)benzonitrile(50 mg, 0.23 mmol) in 1,2-dichloroethane (1 mL) was added 1M BBr₃solution in DCM (0.69 mL, 0.69 mmol) dropwise. The resulting reactionmixture was stirred under microwave irradiation five times (setparameters: T=100° C., t=1 hour) adding further 1M BBr₃ solution in DCM(1 mL) each time. The total amount of 1M BBr₃ solution in DCM used was4.69 mL.

Reaction B: In a vial were added4-Methoxy-2-(trifluoromethoxy)benzonitrile (750 mg, 3.45 mmol),1,2-dichloroethane (5 mL) and then 1M BBr₃ solution in DCM (10.36 mL,10.36 mmol) dropwise. The resulting reaction mixture was stirred undermicrowave irradiation for 1 hour (set T=100° C.). To the reactionmixture further 1M BBr₃ solution in DCM (1 mL) was added and theresulting reaction mixture was stirred under microwave irradiation threemore times (set parameters: T=100° C., t=1.5 hours), adding further 1MBBr₃ solution in DCM (0.8 mL) each time. The total amount of 1M BBr₃solution in DCM used was 13.76 mL.

The two reactions mixtures A and B were added dropwise to a NaHCO₃saturated aqueous solution and the pH was adjusted to 7 with theaddition of solid NaHCO₃. The two phases were separated and the aqueousphase was extracted with DCM (1×) and with EtOAc (2×). The combinedorganic phases were dried and evaporated to dryness to give the titlecompound in mixture with unreacted starting material (1.48 g) as a blackoil. This mixture was used in the next step without furtherpurification.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 11.35 (1H, s), 7.82 (1H, d), 6.91-6.98(2H, m); UPLC_ipqc: 0.88 min, 204 [M+H]+, 202 [M−H]−.

Reference Intermediate R134-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile

A mixture of 2-chloro-5-nitropyridine (70 mg, 0.44 mmol),4-hydroxy-3-(trifluoromethyl)benzonitrile (91 mg, 0.49 mmol), K₂CO₃ (92mg, 0.66 mmol) in DMF (2 mL) was stirred at 50° C. overnight. Water (4mL) was added and a precipitate was formed. The solid was filtered-offand it was triturated with MeOH to give the title compound (85 mg) as abrownish solid.

¹H NMR (400 MHz, CDCl₃): δ ppm 8.99 (1H, d), 8.60 (1H, dd), 8.07 (1H,s), 7.95 (1H, d), 7.48 (1H, d), 7.19-7.32 (1H, m); UPLC_ipqc: 1.1 min,310 [M+H]+.

The following compounds were prepared using the foregoing methodology,reacting the appropriate halo nitroaryl such as2-chloro-5-nitropyridine, 2-chloro-5-nitropyrimidine,1-fluoro-4-nitrobenzene etc. with the appropriately substituted phenolat a suitable temperature, optionally under microwave irradiation. Somefinal products were purified by flash-chromatography (Silica;Cyclohexane/EtOAc or other appropriate solvent system).

R14

3-bromo-4- [(5-nitro-2- pyridinyl) oxy] benzo- nitrile 2- chloro- 5-nitro- pyridine 3-bromo-4- hydroxy- benzo- nitrile ¹H-NMR (400 MHz,CDCl₃): δ ppm 9.01 (1 H, br. s), 8.65-8.56 (1 H, m), 8.02 (1H, s), 7.75(1 H, d), 7.38 (1H, d), 7.25 (1H, d) 1.08 min, 320 [M]+, Br pattern R15

3-(1,1- dimethyl- ethyl)-4- [(5-nitro- 2-pyridinyl) oxy]benzo- nitrile2- chloro- 5- nitro- pyridine 3-(1,1- dimethyl- ethyl)-4- hydroxy-benzo- nitrile (Reference Inter- mediate R10) ¹H NMR (400 MHz, DMSO-d₆):δ ppm 9.09- 9.04 (1H, m), 8.74-8.64 (1H, m), 7.87 (1H, br. s), 7.83-7.77 (1H, m), 7.44 (1H, d), 7.33 (1H, d), 1.32 (9H, s) 1.23 min, 298[M + H]⁺ R16

2-iodo-4- [(5-nitro-2- pyridinyl) oxy]benzo- nitrile 2- chloro- 5-nitro- pyridine 4-hydroxy- 2-iodo- benzo- nitrile (Reference Inter-mediate 11) 1H NMR (400 MHz, DMSO-d₆): δ ppm 9.08 (1H, d), 8.70 (1H,dd), 8.03 (1H, d), 7.98 (1H, d), 7.52 (1H, dd), 7.41 (1H, d) 1.10 minR17

4-[(5-nitro- pyridinyl) oxy]-2- [(trifluoro methyl) oxy]benzo- nitrile2- chloro- 5- nitro- pyridine 4-hydroxy- 2- [(trifluoro- methyl)oxy]benzo- nitrile (Reference Inter- mediate 12) ¹H NMR (400 MHz,CDCl₃): δ ppm 9.02- 9.11 (1H, m), 8.55-8.65 (1H, m), 7.82 (1 H, d),7.25-7.35 (2 H, m), 7.20 (1 H, d) 1.14 min, 326 [M + H]+

Reference Intermediate R182-cyclopropyl-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

Preparation of organometallic solution: to a solution of 0.5M ZnCl₂ inTHF (9 mL) a solution of 0.5M Cyclopropyl Magnesium bromide in THF (9mL) was slowly added at r.t. and the reaction mixture was stirred for 20minutes at r.t.

To a solution of 2-iodo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile(Reference Intermediate R16, 550 mg) and Pd(tBu₃P)₂ (76 mg, 0.15 mmol),warmed at 60° C., were added 6 mL of the organometallic solutionpreviously formed and the reaction mixture was stirred for 1 hour at 60°C. Further 6 mL of the organometallic solution were added and thereaction mixture was stirred for additional 1 hour at 60° C. Further 6mL of the organometallic solution were added and the reaction mixturewas stirred for additional 1 hour at 60° C. After cooling the reactionwas quenched with water (1 mL), diluted with an aqueous saturatedsolution of ammonium chloride (20 mL) and extracted with ethyl acetate(2×50 mL). The organic layer was washed with brine (2×20 mL), dried(Na₂SO₄), filtered and evaporated and the residue was purified by flashchromatography on silica gel (SNAP 50 g), eluting from 100:0 to 80:20n-hexane/ethyl acetate affording the title compound (400 mg) as whitesolid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.06 (1H, d), 8.67 (1H, dd), 7.88 (1H,d), 7.35 (1H, d), 7.23 (1H, dd), 7.01 (1H, dd), 2.17-2.27 (1H, m),1.10-1.19 (2H, m), 0.82-0.90 (2H, m); UPLC_ipqc: 1.13 min, 282 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing Cyclopropyl Magnesium bromide with the appropriate Grignardreagent to form the organozinc reagen.

R19

2-ethyl-4-[(5- nitro-2- pyridinyl) oxy]benzo- nitrile Ethyl magnesiumbromide ¹H-NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, d) 8.56 (1H, dd) 7.72(1H, d) 7.18 (1H, d) 7.10-7.17 (2H, m) 2.95 (2H, q) 1.35 (3H, t) 1.12min, 270 [M + H]+

Reference Intermediate R203-cyclopropyl-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

In a vial 3-bromo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile (ReferenceIntermediate R14, 800 mg) was dissolved in 16.0 mL of toluene.Cyclopropylboronic acid (1073.8 mg, 12.5 mmol) was added, followed byPd(OAc)₂ (56.1 mg, 0.25 mmol) and (Cy)₃P (70.0 mg 0.25 mmol). Then, anaqueous solution (8.0 mL of water) of K₃PO₄ (1855.0 mg, 8.75 mmol) wasadded. The reaction mixture was heated at 80° C. overnight. Aftercooling down to r.t., the mixture was partitioned between brine andEtOAc and the separated aqueous phase was extracted with EtOAc (threetimes). The collected organic were dried over Na₂SO₄, filtered andevaporated. The crude obtained was charged on a silica gel column andeluted with Cyclohexane/EtOAc (from 100:0 to 80:20 Cyclohexane/EtOAc)affording 634 mg of the title compound.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.04 (1H, br. s), 8.69 (1H, dd), 7.75(1H, d), 7.58 (1H, s), 7.41 (2H, t), 1.90-1.80 (1H, m), 0.90-0.73 (4H,m); UPLC_ipqc: 1.12 min, 282 [M+H]⁺.

Reference Intermediate R212-(1-methylethenyl)-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

To a solution of 2-iodo-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile(Reference Intermediate R16, 5.0 g) in DMF (50 mL) were added K₃PO₄(5.77 g, 27.24 mmol), Pd(tBu₃)₂ (696 mg, 1.36 mmol) and4,4,5,5-tetramethyl-2-(1-methylethenyl)-1,3,2-dioxaborolane (3.84 mL,20.43 mmol) and the reaction mixture was stirred for 4 hours at 110° C.After cooling the reaction was diluted with water (100 mL) and extractedwith ethyl acetate (3×100 mL). The organic layer was washed with icecold brine (3×50 mL), dried (Na₂SO₄), filtered and evaporated and theresidue was purified by flash chromatography on silica gel (SNAP 100 g)eluting from 100:0 to 80:20 cyclohexane/ethyl acetate to afford thetitle compound (1.8 g) as white solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 9.08 (1H, d), 8.69 (1H, dd), 7.97 (1H,d), 7.47 (1H, d), 7.40 (2H, d), 5.46 (1H, s), 5.32 (1H, s), 2.16 (3H,s); UPLC_ipqc: 1.14 min, 282 [M+H]+.

Reference Intermediate R222-[(1-methylethyl)oxy]-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile

In a vial 2,4-dihydroxybenzonitrile (300 mg, 2.2 mmol),2-chloro-5-nitropyridine (351.96 mg, 2.22 mmol) and K₂CO₃ (920 mg, 6.62mmol) were dissolved in DMF (5 mL). The reaction was heated for 1 hourunder microwave irradiations (Set Temperature: 110° C.). The reactionmixture was diluted with Et₂O and water, acidified with aqueous 1N HCluntil pH=2, the phases were separated and the organics were dried overNa₂SO₄. The solid was filtered out and the solvent was removed affordingcrude 2-hydroxy-4-[(5-nitro-2-pyridinyl)oxy]benzonitrile (664 mg) as abrown solid. To a solution of this crude in dry DMF (5 mL) potassiumcarbonate (460 mg, 3.33 mmol) and isopropyl bromide (313 μL, 3.33 mmol)were added and the reaction mixture was stirred overnight at 50° C. Thereaction was diluted with brine (10 mL) and extracted with ethyl acetate(2×20 mL). The organic layer was dried (Na₂SO₄), filtered and evaporatedand the residue was purified by flash chromatography on silica gel (SNAP25 g) eluting from 100:0 to 75:25 cyclohexane/ethyl acetate affordingthe title compound (260 mg) as white solid.

¹H-NMR (400 MHz, CDCl₃): δ ppm 9.06 (1H, d), 8.56 (1H, dd), 7.61-7.67(1H, m), 7.15 (1H, d), 6.76-6.84 (2H, m), 4.56-4.68 (1H, m), 1.44 (6H,d).

Reference Intermediate R234-[(5-amino-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile

To a solution of4-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile (ReferenceIntermediate R13, 83 mg) in THF (3 mL)/water (1.5 mL) was added at roomtemperature, iron (75 mg, 1.34 mmol) and NH₄Cl (72 mg, 1.34 mmol) andthe resulting reaction mixture was stirred overnight. The mixture wasfiltered through a small pad of celite washing with EtOAc and water. Tothe filtered mixture was added an aqueous NaHCO₃ saturated solution andthe two phases were separated. The aqueous phase was extracted withEtOAc and the combined organic phases were dried and evaporated todryness. The crude was purified by flash chromatography (companionsystem, 2×12 g Si cartridge, from 100:0 to 70:30 Cyclohexane/EtOAc) toafford the title compound (72 mg).

¹H NMR (400 MHz, CDCl₃): δ ppm 7.97 (1H, s), 7.69-7.79 (2H, m), 7.23(1H, d), 7.16 (1H, dd), 6.93 (1H, d); UPLC_ipqc: 0.91 min, 280 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacing 4-[(5-nitro-2-pyridinyl)oxy]-3-(trifluoromethyl)benzonitrile(Reference Intermediate R13) with the appropriate nitro derivative. Somefinal products were purified by flash-chromatography (Silica or NHcartridge; Cyclohexane/EtOAc or other appropriate solvent system). Insome cases purification by SCX (MeOH and then 2M ammonia solution inMeOH) was run before the usual flash-chromatography.

R24

4-[(5- amino- 2- pyridinyl) oxy]-3- cyclo- propyl- benzo- nitrile3-cyclo- propyl-4- [(5-nitro- 2-pyridinyl) oxy]benzo- nitrile (ReferenceInter- mediate R20) ¹H NMR (400 MHz, DMSO-d6): δ ppm 7.61- 7.52 (2H, m),7.42 (1H, s), 7.16-7.09 (1H, m), 6.89 (2H, t), 5.19 (2H, br. s),2.14-2.04 (1H, m), 0.97- 0.89 (2H, m), 0.82- 0.75 (2H, m) 0.86 min, 252[M + H]+ R25

4-[(5- amino- 2- pyridinyl) oxy]-3-(1,1- dimethyl- ethyl) benzo- nitrile3-(1,1- dimethyl- ethyl)-4- [(5-nitro-2- pyridinyl) oxy]benzo- nitrile(Reference Inter- mediate R15) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 7.72(1H, br. s), 7.65-7.57 (2H, m), 7.16-7.08 (1H, m), 6.89- 6.78 (2H, m),5.28-5.19 (2H, m), 1.39 (9H, s) 1.02 min, 268 [M + H]⁺ R26

4-[(5-amino- 2-pyridinyl) oxy]-2-(1- methyl- ethenyl) benzo- nitrile2-(1- methyl- ethenyl)-4- [(5-nitro-2- pyridinyl) oxy]benzo- nitrile(Reference Inter- mediate R21) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 7.79(1H, d), 7.62 (1H, d), 7.12 (1H, dd), 7.05 (1H, d), 6.98 (1H, dd), 6.89(1H, d), 5.40 (1H, s), 5.28 (2H, br. s.), 5.23 (1H, s), 2.12 (3H, s)0.90 min, 252 [M + H]+ R27

4-[(5-amino- 2- pyridinyl) oxy]-2- ethyl- benzo- nitrile 2-ethyl-4-[(5-nitro- 2-pyridinyl) oxy]benzo- nitrile (Reference Inter- mediateR19) ¹H-NMR (400 MHz, CDCl3): δ ppm 7.77 (1H, d) 7.58 (1H, d) 7.15 (1H,dd) 7.00 (1H, d) 6.92 (1H, dd) 6.86(1H, d) 3.62 (2H, br. s.) 2.86 (2H,q) 1.29 (3H, t) 0.86 min, 240 [M + H]+ R28

4-[(5-amino- 2-pyridinyl) oxy]methyl- ethyl)oxy] benzo- nitrile 2-[(1-methyl- ethyl)oxy]- 4-[(5-nitro- 2-pyridinyl) oxy]benzo- nitrile(Reference Inter- mediate R22) ¹H-NMR (400 MHz, DMSO-d6): δ ppm 7.63(1H, d), 7.61 (1H, d), 7.12 (1H, dd), 6.88 (1H, d), 6.84 (1H, d), 6.51(1H, dd), 5.29 (2H, br. s.), 4.66-4.77 (1H, m), 1.29 (6H, d) 0.89 min,270 [M + H]+ R29

4-[(5-amino- 2-pyridinyl) oxy]-2- [(trifluoro- methyl) oxy]benzo-nitrile 4-[(5- nitro-2- pyridinyl) oxy]-2- [(trifluoro- methyl)oxy]benzo- nitrile (Reference Inter- mediate R16) ¹H NMR (400 MHz,CDCl₃): δ ppm 7.77 (1H, d), 7.66 (1H, d), 7.17 (1 H, dd), 7.11 (1 H, s),7.06 (1 H, dd), 6.89 (1 H, d) 0.94 min, 296 [M + H]+

Reference Intermediate R30 1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate

(2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acid (121.4 mg,0.60 mmol) was dissolved in N,N-Dimethylformamide (1 mL).N,N-Diisopropylethylamine (0.126 mL, 0.72 mmol) andO-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (227.2 mg, 0.60 mmol) were added. The reactionmixture was stirred at r.t. for 30 min.4-[(5-amino-2-pyridinyl)oxy]-3-cyclopropylbenzonitrile (ReferenceIntermediate R24, 100 mg) was dissolved in 1.0 mL of DMF and theobtained solution was added to the reaction mixture. The reactionmixture was stirred and heated at 60° C. for 2 h. After cooling down tor.t., the reaction mixture was evaporated under vacuum and the crudeobtained was charged on a silica gel column and eluted withCyclohexane/EtOAc (from 100:0 to 50:50 Cyclohexane/EtOAc, then plateauat 50:50) affording 133 mg of the title compound.

¹H-NMR (400 MHz, CDCl₃): δ ppm 8.42 (1H, br. s), 8.20-8.10 (2H, m),7.51-7.44 (1H, m), 7.32-7.23 (1H, m), 7.08 (1H, d), 7.03-6.95 (1H, m),4.95 (1H, br. s), 4.16-4.05 (1H, m), 2.07-1.95 (2H, m), 1.77-1.68 (1H,m), 1.47 (9H, s), 1.04 (3H, t), 0.95-0.88 (2H, m), 0.71-0.64 (2H, m);UPLC_ipqc: 1.14 min, 437 [M+H]⁺.

The following compounds were prepared using the foregoing methodology,replacing (2R)-2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)butanoic acidwith the appropriate aminoacid and4-[(5-amino-2-pyridinyl)oxy]-3-cyclopropylbenzonitrile (Intermediate165) with the appropriate anilin. The reaction was carried out at asuitable temperature ranging from r.t. to high temperature. Finalproducts were purified by flash-chromatography (Silica;Cyclohexane/EtOAc or other appropriate solvent system).

R31

1,1- dimethyl- ethyl ((1R)-1- {[(6-{[4- cyano-2- (1,1- dimethyl- ethyl)phenyl] oxy}-3- pyri- dinyl) amino] carbonyl} propyl) carbamate (2R)-2-({[(1,1- dimethyl- ethyl) oxy] carbonyl} amino) butanoic acid 4-[(5-amino-2- pyri- dinyl) oxy]-3- (1,1- dimethyl- ethyl) benzo- nitrile(Refer- ence Inter- mediate R25) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 10.20(1H, br. s), 8.38 (1H, br. s), 8.20- 8.11 (1H, m), 7.77 (1H, br. s),7.70- 7.64 (1H, m), 7.15 (1H, d), 7.05 (2H, d), 4.04-3.91 (1H, m),1.77-1.55 (2H, m), 1.41- 1.34 (18H, m), 0.93-0.87 (3H, m) 1.25 min, 453[M + H]⁺. R32

1,1- dimethyl- ethyl ((1R)-1- {[(6-{[4- cyano-3- (1-methyl- ethenyl)phenyl] oxy}-3- pyri- dinyl) amino] carbonyl} propyl) carbamate (2R)-2-({[(1,1- dimethyl- ethyl) oxy] carbonyl} amino) butanoic acid 4-[(5-amino-2- pyri- dinyl) oxy]-2- (1- methyl- ethenyl) benzo- nitrile(Refer- ence Inter- mediate R26) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.21(1 H, br. s.), 8.39-8.47 (1 H, m), 8.17 (1 H, dd), 7.86 (1 H, d), 7.24(1 H, d), 7.17 (2 H, d), 7.01-7.10 (1 H,m), 5.43 (1 H, s), 5.27 (1 H,s), 3.95- 4.05 (1 H, m), 2.14 (3 H, s), 1.57-1.79 (2 H, m), 1.40 (9 H,s), 0.92 (3 H, t) 1.16 min, 437 [M + H]+ R33

1,1- dimethyl- ethyl {(1R)-1- [({6-[(4- cyano-3- ethyl- phenyl) oxy]-3-pyri- dinyl} amino) carbonyl] propyl} carbamate (2R)-2- ({[(1,1-dimethyl- ethyl) oxy] carbonyl} amino) butanoic acid 4-[(5- amino-2-pyri- dinyl) oxy]-2- ethyl- benzo- nitrile (Refer- ence Inter- mediateR27) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.25 (1H, s) 8.43 (1H, d) 8.16(1H, dd) 7.80 (1H, d) 7.20 (1H, d) 7.15 (1H, d) 7.05- 7.11 (2H, m)3.94-4.02 (1H, m) 2.79 (2H, q) 1.53- 1.76 (2H, m) 1.39 (9H, s) 1.21 (3H,t) 0.91 (3H, t) R34

1,1- dimethyl- ethyl [(1R)-1- ({[6-({4- cyano-3- [(1- methyl- ethyl)oxy]phenyl} oxy)-3- pyri- dinyl] amino} carbonyl) propyl] carbamate (2R)-2-({[(1,1- dimethyl- ethyl) oxy] carbonyl} amino) butanoic acid 4-[(5-amino-2- pyri- dinyl) oxy]-2- [(1- methyl- ethyl)oxy] benzo- nitrile(Refer- ence Inter- mediate R28) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.24(1H, br. s.), 8.42 (1H, d), 8.16 (1H, dd), 7.70 (1H, d), 7.15 (1H, d),7.09 (1H, d), 7.02 (1H, d), 6.72 (1H, dd), 4.70-4.81 (1H, m), 3.94-4.02(1H, m), 1.54- 1.77 (2H, m), 1.39 (9H, s), 1.30 (6H, d), 0.91 (3H, t)1.15 min, 455 [M + H]+ R35

1,1- dimethyl- ethyl [(1R)-1- ({[6-({4- cyano-3- [(tri- fluoro- methyl)oxy] phenyl} oxy)-3- pyri- dinyl] amino} carbonyl) propyl] carbamate(2R)-2- ({[(1,1- dimethyl- ethyl) oxy] carbonyl} amino) butanoic acid4-[(5- amino-2- pyri- dinyl) oxy]-2- [(tri- fluoro- methyl) oxy] benzo-nitrile (Refer- ence Inter- mediate R29) ¹H NMR (400 MHz, CDCl₃): δ ppm8.64 (1H, br. s.), 8.26 (1H, d), 8.15-8.23 (1H, m), 7.70 (1H, d), 7.20(1H, s), 7.12- 7.18 (1H, m), 7.02 (1H, d), 4.93-5.06 (1H, m), 4.10- 4.21(1H, m), 1.93-2.12 (1H, m), 1.67-1.83 (1H, m), 1.49 (9H, s), 1.06 (3H,t) 1.18 min, 481 [M + H]+, 479 [M − H ]−

Reference Intermediate R361,1-dimethylethyl((1R)-1-{[(6-{[4-cyano-3-(1-methylethyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate

To a solution of1,1-dimethylethyl((1R)-1-{[(6-{[4-cyano-3-(1-methylethenyl)phenyl]oxy}-3-pyridinyl)amino]carbonyl}propyl)carbamate(Reference Intermediate R32, 73 mg) in MeOH (10 mL) was added Pd 10% w/won activated carbon (14 mg) and the reaction mixture was stirred for 30minutes under H₂ atmosphere (P=1 atm). The catalyst was filtered off andthe solvent removed under reduced pressure. The residue was purified byflash chromatography on silica gel (SNAP 10 g) eluting from 75:25 to40:60 cyclohexane/ethyl acetate affording the title compound (62 mg) aswhite solid.

¹H-NMR (400 MHz, DMSO-d₆): δ ppm 10.24 (1H, br. s.), 8.42 (1H, d), 8.16(1H, dd), 7.78 (1H, d), 7.24 (1H, d), 7.15 (1H, d), 7.07-7.11 (1H, m),7.05 (1H, dd), 3.95-4.02 (1H, m), 3.19-3.27 (1H, m), 1.57-1.76 (2H, m),1.39 (9H, s), 1.26 (6H, d), 0.91 (3H, t); UPLC_ipqc: 1.20 min, 439[M+H]+.

Reference Intermediate R37(2R)-2-amino-N-{6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}butanamide

1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Reference Intermediate R30, 133 mg) was dissolved in DCM (6 mL) and, at0° C., TFA (3.0 mL) was slowly added. The reaction mixture was stirredat that temperature for 2 h. After the removal of the volatiles, thecrude obtained was charged on a SCX cartridge and eluted with MeOH andthen 2M NH₃ in MeOH affording 102 mg of the title compound.

¹H-NMR (400 MHz, CDCl₃): δ ppm 9.68 (1H, br. s), 8.32-8.18 (2H, m),7.51-7.43 (1H, m), 7.25-7.31 (1H, m), 7.08 (1H, d), 6.99 (1H, d),3.59-3.51 (1H, m), 2.06-1.95 (2H, m), 1.73-1.63 (1H, m), 1.03 (3H, t),0.95-0.89 (2H, m), 0.74-0.63 (2H, m); UPLC_ipqc: 0.68 min, 337 [M+H]⁺.

The following compounds were prepared using the foregoing methodology,replacing 1,1-dimethylethyl{(1R)-1-[({6-[(4-cyano-2-cyclopropylphenyl)oxy]-3-pyridinyl}amino)carbonyl]propyl}carbamate(Reference Intermediate R30) with the appropriate N—BOC protected amine.Final products were purified by SCX (MeOH and then 2M ammonia solutionin MeOH) and fractions eluted with ammonia, containing the product, wereconcentrated to provide the free-base. Alternatively, after removing thevolatiles, to the crude taken up with an appropriate organic solvent wasadded NaHCO₃ saturated aqueous solution, the two phases were separatedand the organic layer was dried, filtered and evaporated affording thefinal compound as the free-base.

R38

(2R)-2- amino-N- (6-{[4- cyano-2- (1,1- dimethyl- ethyl) phenyl] oxy}-3-pyridinyl) butan- amide 1,1-dimethyl- ethyl ((1R)- 1-{[(6-{[4-cyano-2-(1,1- dimethyl- ethyl)phenyl] oxy}-3- pyridinyl) amino]carbonyl} propyl) carbamate (Reference Inter- mediate R31) ¹H NMR (400MHz, DMSO-d₆): δ ppm 8.43 (1H, br. s), 8.25-8.15 (1H, m), 7.78 (1H, br.s), 7.70-7.65 (1H, m), 7.14 (1H, d), 7.05 (1H, d), 3.20- 3.15 (1H, m),1.74-1.61 (1H, m), 1.57- 1.45 (1H, m), 1.36 (9H, s), 0.93 (3H, t) 0.79min, 353 [M + H]⁺ R39

(2R)-2- amino-N- (6-{[4- cyano-3- (1-methyl- ethyl) phenyl] oxy}-3-pyridinyl) butan- amide 1,1-dimethyl- ethyl ((1R)- 1-{[(6-{[4-cyano-3-(1- methylethyl) phenyl]oxy}- 3-pyridinyl) amino] carbonyl}propyl) carbamate (Reference Inter- mediate R36) ¹H-NMR (400 MHz,DMSO-d₆): δ ppm 8.48 (1H, d) 8.22 (1H, dd) 7.79 (1H, d) 7.24 (1H, d)7.15 (1H, d) 7.05 (1H, dd) 3.19- 3.30 (2H, m) 1.61-1.74 (1H, m) 1.45-1.56 (1H, m) 1.26 (6H, d) 0.91 (3H, t) 0.76 min, 339 [M + H]+ R40

(2R)-2- amino-N- {6-[(4- cyano-3- ethyl- phenyl) oxy]-3- pyridinyl}butan- amide 1,1-dimethyl- ethyl {(1R)- 1-[({6-[(4- cyano-3-ethylphenyl) oxy]-3- pyridinyl} amino) carbonyl] propyl} carbamate(Reference Inter- mediate R33) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.48(1H, d) 8.22 (1H, dd) 7.80 (1H, d) 7.19 (1H, d) 7.15 (1H, d) 7.07 (1H,dd) 3.22- 3.29 (1H, m) 2.79 (2H, q) 1.60-1.74 (1H, m) 1.44- 1.56 (1H, m)1.21 (3H, t) 0.91 (3H, t) R41

(2R)-2- amino-N- [6-({4- cyano-3- [(1-methyl- ethyl)oxy] phenyl} oxy)-3-pyridinyl] butan- amide 1,1-dimethyl- ethyl [(1R)- 1-({[6-({4- cyano-3-[(1-methyl- ethyl)oxy] phenyl} oxy)-3- pyridinyl] amino} carbonyl)propyl] carbamate (Reference Inter- mediate R34) ¹H-NMR (400 MHz,CDCl₃): δ ppm 9.70 (1H, br. s.), 8.26-8.38 (2H, m), 7.54 (1H, d), 7.01(1H, d), 6.74 (1H, d), 6.68 (1H, dd), 4.52- 4.66 (1H, m), 3.45-3.54 (1H,m), 1.97- 2.10 (1H, m), 1.65-1.76 (1H, m), 1.41 (6H, d), 1.06 (3H, t)R42

(2R)-2- amino-N- [6-({4- cyano-3- [(trifluoro- methyl) oxy] phenyl}oxy)-3- pyridinyl] butan- amide 1,1-dimethyl- ethyl [(1R)- 1-({[6-({4-cyano-3- [(trifluoro- methyl)oxy] phenyl} oxy)-3- pyridinyl] amino}carbonyl) propyl] carbamate (Reference Inter- mediate R35) ¹H NMR (400MHz, CDCl₃): δ ppm 9.76 (1H, s), 8.25- 8.43 (2H, m), 7.70 (1H, d), 7.20(1H, s), 7.16 (1H, dd), 7.06 (1H, d), 3.44-3.59 (1H, m), 1.53- 2.12 (2H,m), 1.07 (3H, t) 0.72 min, 381 [M + H]+, 379 [M − H]−.

Reference Example RE1(5R)-5-ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

Method A

To a solution of(2R)-2-amino-N-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)butanamide(Reference Intermediate R6, 120 mg) in dry dichloromethane (8 mL) TEA(0.265 mL, 1.903 mmol) was added and the reaction mixture was cooled to0° C. A solution of triphosgene (50.8 mg, 0.171 mmol) in drydichloromethane (DCM) (2 mL) was slowly added and the reaction mixturewas stirred for 30 minutes at the same temperature. The reaction wasquenched with water (2 mL) and two phases were separated. The organiclayer was dried over sodium sulphate, filtered and evaporated and theresidue was purified by silica gel chromatography (Biotage system, 10 gSNAP column) with as eluent a gradient cyclohexane/ethyl acetate 80/20to cyclohexane/ethyl acetate 50/50 to afford the title compound as awhite solid (108 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.61 (1H, s), 8.12 (1H, d), 7.82 (1H,dd), 7.17 (1H, d), 7.08 (1H, d), 6.79 (1H, d), 6.63 (1H, dd), 4.25-4.18(1H, m), 3.77 (3H, s), 2.15 (3H, s), 1.89-1.62 (2H, m), 0.95 (3H, t):UPLC_B: 0.79 min, 342 [M+H]+.

Reference RE2(5R)-5-ethyl-5-methyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-2,4-imidazolidinedione

To a solution ofN1-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}-3-pyridinyl)-D-isovalinamide(Reference Intermediate R8 42 mg) in dry dichloromethane (6 mL), TEA(0.089 mL, 0.638 mmol) was added. The mixture was cooled down to 0° C.and a solution of triphosgene (17.03 mg, 0.057 mmol) in drydichloromethane (1.500 mL) was added dropwise. The mixture was stirredat that temperature for 1 hour, then a solution of triphosgene (17.03mg, 0.057 mmol) in dry dichloromethane (DCM) (1.500 mL) was addeddropwise again. The reaction was stirred for 30 minutes, it wasmaintained in the ice-bath and quenched with water (10 mL). The mixturewas allowed to reach the room temperature then it was extracted withdichloromethane (3×7 mL). The combined organic layers were dried oversodium sulphate, filtered and evaporated. The residue obtained waspurified by flash chromatography on silica gel using a 10 g SNAP columnand cyclohexane/ethyl acetate as eluents from 80/20 to 50/50 (Biotagesystem). This afforded the title compound as a white solid (24 mg).

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.57 (1H, s), 8.13 (1H, d), 7.83 (1H,dd), 7.17 (1H, d), 7.07 (1H, d), 6.79 (1H, d), 6.62 (1H, dd), 3.76 (3H,s), 2.14 (3H, s), 1.57-1.86 (2H, m), 1.39 (3H, s), 0.86 (3H, t); UPLC_B:0.83 min, 354 [M−H]+.

Method for Reference Examples RE3 to RE84-{[4-(4,4-dimethyl-2,5-dioxo-1-imidazolidinyl)phenyl]oxy}-2-(methyloxy)benzonitrile

N¹-(4-{[4-cyano-3-(methyloxy)phenyl]oxy}phenyl)-2-methylalaninamide(77.0 mg) was dissolved in DCM (10 mL). Triethylamine (0.218 mL, 1.57mmol) was added and the obtained mixture was cooled at 0° C.Bis(trichloromethyl)carbonate (68.1 mg, 0.22 mmol) was dissolved in 5 mLof DCM and the obtained solution was added dropwise to the reactionmixture. The reaction mixture was stirred at 0° C. After 15 min, thereaction mixture was evaporated in vacuo to obtain the crude productthat was purified by silica gel chromatography (from 100:0 to 50:50Cyclohexane/EtOAc in 10 CV; then 50:50 Cyclohexane/EtOAc for 10 CV) toobtain 65.1 mg of the title compound as a white solid.

¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.56 (1H, br. s.) 7.72 (1H, d)7.42-7.49 (2H, m) 7.19-7.29 (2H, m) 6.97 (1H, d) 6.57 (1H, dd) 3.89 (3H,s) 1.41 (6H, s); UPLC_ipqc: 0.93 min, 352 [M+H]+.

The following compounds were prepared using the foregoing methodology,replacingN¹-(4-{[4-cyano-3-(methyloxy)phenyl]oxy}phenyl)-2-methylalaninamide withthe appropriate amine. Final products were purified byflash-chromatography (Silica cartridge; Cyclohexane/EtOAc or otherappropriate solvent system).

RE3

3-(1,1- dimethyl- ethyl)-4- ({5-[(4R)- 4-ethyl- 2,5-dioxo- 1- imidazol-idinyl]-2- pyridinyl} oxy) benzo- nitrile (2R)-2- amino-N- (6-{[4-cyano-2- (1,1-dimethyl- ethyl)phenyl] oxy}-3- pyridinyl) butan- amide(Reference Inter- mediate R38) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.64(1H, br. s), 8.19 (1H, br. s), 7.96-7.90 (1H, m),7.82 (1H, br. s),7.76-7.68 (1H, m), 7.30 (1H, d), 7.19 (1H, d), 4.25- 4.17 (1H, m),1.86-1.77 (1H, m), 1.76- 1.66 (1H, m), 1.35 (9H, s), 0.95 (3H, t) RE4

4-({5- [(4R)-4- ethyl-2,5- dioxo-1- imidazol- idinyl]-2- pyridinyl}oxy)-2-(1- methyl- ethyl) benzo- nitrile (2R)-2- amino-N- (6-{[4-cyano-3- (1-methyl- ethyl)phenyl] oxy}-3- pyridinyl) butan- amide(Reference Inter- mediate R39) ¹H-NMR (400 MHz, DMSO-d6): δ ppm 8.66(1H, s) 8.19 (1H, d) 7.93 (1H, dd) 7.84 (1H, d) 7.36 (1H, d) 7.28 (1H,d) 7.18 (1H, dd) 4.19- 4.25 (1H, m) 3.21-3.30 (1H, m) 1.77- 1.87 (1H, m)1.65-1.76 (1H, m) 1.27 (6H, d) 0.96 (3H, t) 1.03 min, 365 [M + H]+ RE5

3-cyclo- propyl-4- ({5-[(4R)- 4-ethyl- 2,5-dioxo- 1-imidazol- idinyl]-2-pyridinyl} oxy) benzo- nitrile (2R)-2- amino-N- {6-[(4- cyano-2- cyclo-propyl- phenyl) oxy]-3- pyridinyl} butan- amide (Reference Inter-mediate R37) ¹H NMR (400 MHz, DMSO-d₆): δ ppm 8.64 (1H, br. s),8.14-8.11 (1H, m), 7.96- 7.84 (1H, m), 7.71-7.66 (1H, m), 7.52 (1H, br.s), 7.29 (2H, d), 4.24-4.18 (1H, m), 1.97- 1.89 (1H, m), 1.86-1.78 (1H,m), 1.75- 1.67 (1H, m), 0.95 (3H, t), 0.91-0.85 (2H, m), 0.81- 0.75 (2H,m) 0.98 min, 363 [M + H]⁺. RE6

4-({5- [(4R)-4- ethyl-2,5- dioxo-1- imidazol- idinyl]-2- pyridinyl}oxy)-2-(1- methyl- ethyl) benzo- nitrile (2R)-2- amino-N- {6-[(4-cyano-3- ethyl- phenyl) oxy]-3- pyridinyl} butan- amide (ReferenceInter- mediate R40) ¹H-NMR (400 MHz, DMSO-d₆): δ ppm 8.66 (1H, s) 8.19(1H, dd) 7.93 (1H, dd) 7.85 (1H, d) 7.32 (1H, d) 7.28 (1H, dd) 7.19 (1H,dd) 4.19- 4.25 (1H, m) 2.82 (2H, q) 1.77-1.88 (1H, m) 1.65- 1.77 (1H, m)1.23 (3H, t) 0.96 (3H, t) 0.98 min, 351 [M + H]+ RE7

4-({5- [(4R)-4- ethyl-2,5- dioxo-1- imidazol- idinyl]-2- pyridinyl}oxy)-2- [(trifluoro- methyl)oxy] benzo- nitrile (2R)-2- amino-N- [6-({4-cyano-3- [(trifluoro- methyl) oxy] phenyl} oxy)-3- pyridinyl] butan-amide (Reference Inter- mediate R42) ¹H NMR (400 MHz, DMSO-d6): δ ppm8.67 (1H, s), 8.19- 8.26 (1H, m), 8.14 (1H, d), 7.94-8.02 (1H, m), 7.66(1H, s), 7.43- 7.51 (1H, m), 7.36 (1H, d), 4.18-4.27 (1H, m), 1.63- 1.91(2H, m), 0.96 (3H, t) 1.02 min, 407 [M + H]+, 405 [M − H]− RE8

4-({5- [(4R)-4- ethyl-2,5- dioxo-1- imidazol- idinyl]-2- pyridinyl}oxy)-2- [(1-methyl- ethyl)oxy] benzo- nitrile (2R)-2- amino-N- [6-({4-cyano-3- [(1-methyl- ethyl)oxy] phenyl} oxy)-3- pyridinyl] butan- amide(Reference Inter- mediate R41) ¹H-NMR (400 MHz, DMSO-d6): δ ppm 8.65(1H, s), 8.20 (1H, d), 7.93 (1H, dd), 7.76 (1H, d), 7.27 (1H, d), 7.16(1H, d), 6.84 (1H, dd), 4.74- 4.85 (1H, m), 4.17-4.26 (1H, m), 1.76-1.89 (1H, m), 1.65-1.76 (1H, m), 1.31 (6H, d), 0.96 (3H,t) 1.00 min, 381[M + H]+.

Biological Example 1

The ability of the compounds of the invention to modulate thevoltage-gated potassium channel subtypes Kv3.2/3.1 may be determinedusing the following assay.

Cell Biology

To assess compound effects on human Kv3.2 channels (hKv3.2), a stablecell line expressing hKv3.2 was created by transfecting Chinese HamsterOvary (CHO)-K1 cells with a pCIH5-hKv3.2 vector. Cells were cultured inDMEM/F12 medium supplemented by 10% Foetal Bovine Serum, 1×non-essential amino acids (Invitrogen) and 500 ug/ml of Hygromycin-B(Invitrogen). Cells were grown and maintained at 37° C. in a humidifiedenvironment containing 5% CO₂ in air.

To assess compound effects on human Kv3.1 channels (hKv3.1),CHO/Gam/E1A-clone22 alias CGE22 cells were transduced using a hKv3.1BacMam reagent. This cell line was designed to be an improvedCHO-K1-based host for enhanced recombinant protein expression ascompared to wild type CHO-K1. The cell line was generated following thetransduction of CHO-K1 cells with a BacMam virus expressing theAdenovirus-Gam1 protein and selection with Geneticin-G418, to generate astable cell line, CHO/Gam-A3. CHO/Gam-A3 cells were transfected withpcDNA3-E1A-Hygro, followed by hygromycin-B selection and FACS sorting toobtain single-cell clones. BacMam-Luciferase and BacMam-GFP viruses werethen used in transient transduction studies to select the clone based onhighest BacMam transduction and recombinant protein expression. CGE22cells were cultured in the same medium used for the hKv3.2 CHO-K1 stablecell line with the addition of 300 ug/ml hygromycin-B and 300 ug/mlG418. All other conditions were identical to those for hKv3.2 CHO-K1cells. The day before an experiment 10 million CGE22 cells were platedin a T175 culture flask and the hKv3.1 BacMam reagent (pFBM/human Kv3.1)was added (MOI of 50). Transduced cells were used 24 hours later.

Cell Preparation for IonWorks Quattro™ Experiments

The day of the experiment, cells were removed from the incubator and theculture medium removed. Cells were washed with 5 ml of Dulbecco's PBS(DPBS) calcium and magnesium free and detached by the addition of 3 mlVersene (Invitrogen, Italy) followed by a brief incubation at 37° C. for5 minutes. The flask was tapped to dislodge cells and 10 ml of DPBScontaining calcium and magnesium was added to prepare a cell suspension.The cell suspension was then placed into a 15 ml centrifuge tube andcentrifuged for 2 min at 1200 rpm. After centrifugation, the supernatantwas removed and the cell pellet re-suspended in 4 ml of DPBS containingcalcium and magnesium using a 5 ml pipette to break up the pellet. Cellsuspension volume was then corrected to give a cell concentration forthe assay of approximately 3 million cells per ml.

All the solutions added to the cells were pre-warmed to 37° C.

Electrophysiology

Experiments were conducted at room temperature using IonWorks Quattro™planar array electrophysiology technology (Molecular Devices Corp.) withPatchPlate™ PPC. Stimulation protocols and data acquisition were carriedout using a microcomputer (Dell Pentium 4). Planar electrode holeresistances (Rp) were determined by applying a 10 mV voltage step acrosseach well. These measurements were performed before cell addition. Aftercell addition and seal formation, a seal test was performed by applyinga voltage step from −80 mV to −70 mV for 160 ms. Following this,amphotericin-B solution was added to the intracellular face of theelectrode to achieve intracellular access. Cells were held at −70 mV.Leak subtraction was conducted in all experiments by applying 50 mshyperpolarizing (10 mV) prepulses to evoke leak currents followed by a20 ms period at the holding potential before test pulses. From theholding potential of −70 mV, a first test pulse to −15 mV was appliedfor 100 ms and following a further 100 ms at −70 mV, a second pulse to40 mV was applied for 50 ms. Cells were then maintained for a further100 ms at −100 mV and then a voltage ramp from −100 mV to 40 mV wasapplied over 200 ms. In all experiments, the test pulses protocol wasperformed in the absence (pre-read) and presence (post-read) of the testcompound. Pre- and post-reads were separated by the compound additionfollowed by a 3 minute incubation.

Solutions and Drugs

The intracellular solution contained the following (in mM): K-gluconate100, KCl 54, MgCl₂ 3.2, HEPES 5, adjusted to pH 7.3 with KOH.Amphotericin-B solution was prepared as 50 mg/ml stock solution in DMSOand diluted to a final working concentration of 0.1 mg/ml inintracellular solution. The external solution was Dulbecco's PhosphateBuffered Saline (DPBS) and contained the following (in mM): CaCl₂ 0.90,KCl 2.67, KH₂PO₄ 1.47, MgCl.6H₂O 0.493, NaCl 136.9, Na₃PO₄ 8.06, with apH of 7.4.

Compounds of the invention (or reference compounds such asN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylureawere dissolved in dimethylsulfoxide (DMSO) at a stock concentration of10 mM. These solutions were further diluted with DMSO using a Biomek FX(Beckman Coulter) in a 384 compound plate. Each dilution (1 μL) wastransferred to another compound plate and external solution containing0.05% pluronic acid (66 μL) was added. 3.5 μL from each plate containinga compound of the invention was added and incubated with the cellsduring the IonWorks Quattro™ experiment. The final assay dilution was200 and the final compound concentrations were in the range 50 μM to 50nM.

Data Analysis

The recordings were analysed and filtered using both seal resistance(>20 MΩ) and peak current amplitude (>500 pA at the voltage step of 40mV) in the absence of compound to eliminate unsuitable cells fromfurther analysis. Kv3 channel-mediated outward currents were measureddetermined from the mean amplitude of the current over the final 10 msof the −15 mV voltage pulse minus the mean baseline current at −70 mVover a 10 ms period just prior to the −15 mV step. This Kv3 channelcurrents following addition of the test compound were then compared withthe currents recorded prior to compound addition. Data were normalisedto the maximum effect of the reference compound (50 microM ofN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylurea)and to the effect of a vehicle control (0.5% DMSO). The normalised datawere analysed using ActivityBase or Excel software. The concentration ofcompound required to increase currents by 50% of the maximum increaseproduced by the reference compound (pEC50) was determined by fitting ofthe concentration-response data using a four parameter logisticfunction.

N-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylureawas obtained from ASINEX (Registry Number: 552311-06-5).

All the Example compounds were tested in the above assay anddemonstrated potentiation of Kv3.1 or Kv3.2 or Kv3.1 and Kv3.2 (hereinafter “Kv3.1 and/or Kv3.2”) whole-cell currents of, on average, at least20% of that observed with 50 microMN-cyclohexyl-N-[(7,8-dimethyl-2-oxo-1,2-dihydro-3-quinolinyl)methyl]-N′-phenylurea.Thus, in the recombinant cell assays of Biological Example 1, all of theExample compounds act as positive modulators. As used herein, a Kv3.1and/or Kv3.2 positive modulator is a compound which has been shown toproduce at least 20% potentiation of whole-cell currents mediated byhuman Kv3.1 and/or human Kv3.2 channels recombinantly expressed inmammalian cells, as determined using the assays described in BiologicalExample 1 (Biological Assays).

A secondary analysis of the data from the assays described in BiologicalExample 1 investigates the effect of the compounds on rate of rise ofthe current from the start of the depolarising voltage pulses. Themagnitude of the effect of a compound can be determined from the timeconstant (Tau_(act)) obtained from a non-linear fit, using the equationgiven below, of the rise in Kv3.1 or Kv3.2 currents following the startof the −15 mV depolarising voltage pulse.

Y=(Y0−Ymax)*exp(−K*X)+Ymax

where:

-   -   Y0 is the current value at the start of the depolarising voltage        pulse;    -   Ymax is the plateau current;    -   K is the rate constant, and Tau_(act) is the activation time        constant, which is the reciprocal of K.

Similarly, the effect of the compounds on the time taken for Kv3.1 andKv3.2 currents to decay on closing of the channels at the end of the −15mV depolarising voltage pulses can also be investigated. In this lattercase, the magnitude of the effect of a compound on channel closing canbe determined from the time constant (Tau_(deact)) of a non-linear fitof the decay of the current (“tail current”) immediately following theend of the depolarising voltage pulse.

The time constant for activation (Tau_(act)) has been determined forseveral of the compounds of the Examples. FIG. 1 shows the data for twocompounds of the invention. Table 1 provides the Tau_(act) data for allof the Examples analysed in this way.

FIG. 1 a shows hKv3.2 currents recorded using the assay described inBiological Example 1. Data shown are the individual currents over theperiod of the depolarising voltage step to −15 mV recorded from 4different cells at two concentrations of compound (Reference ExampleRE1). The data are fitted by a single exponential curve (solid lines)using the fitting procedure in Prism version 5 (Graphpad Software Inc).

FIG. 1 b shows hKv3.2 currents recorded using the assay described inBiological Example 1. Data shown are the individual currents over theperiod of the depolarising voltage step to −15 mV recorded from 2different cells at two concentrations of the compound of ReferenceExample RE3. The data are fitted by a single exponential curve (solidlines) using the fitting procedure in Prism version 5 (Graphpad SoftwareInc).

TABLE 1 Summary hKv3.2 data from the analysis of activation time(Tau_(act)). To allow for comparison between compounds, the compoundconcentration chosen was that which produced a similar current(~0.3nA)at the end of the voltage pulse, with the exception of thevehicle, where maximum currents were <0.1nA. Number of ExampleConcentration (μM) Tau_(act) mean (ms) Standard Deviation experimentsVehicle — 7.1 1.7 6 (cells) RE1 6.25 9.9 2.2 5 RE2 12.5 7.3 1.8 4Example 15 0.2 50.1 7.5 5 Example 16 0.4 19.3 1.0 4 Example 25 6.25 7.873.24 4 RE3 0.2 23.0 6.2 4 RE4 0.8 9.2 2.3 2 RE5 3.1 13.0 2.3 2 RE6 3.18.2 2.0 2 RE7 3.1 10.4 2.8 2 RE8 3.1 9.7 1.0 2 Example 65 0.8 24.0 3.6 2Example 62 0.4 34.8 4.9 2 Example 61 0.8 31.5 4.0 2 Example 51 1.6 21.30.1 2 Example 54 1.6 14.8 1.9 2 Example 63 0.4 28.0 0.4 2 Example 64 1.625.0 2.1 2

As can be seen from Table 1, in the absence of compound and presence ofvehicle the Tau_(act) was 7.1±1.7 msec. A range of Tau_(act) values(7.3-50.1 msec) was observed in the presence of the test compounds wheneach was tested at a concentration that increased the Kv3.2 current to asimilar level (˜0.3 nA).

Kv3.1 and Kv3.2 channels must activate and deactivate very rapidly inorder to allow neurons to fire actions potentials at high frequency(Rudy and McBain, 2001, Trends in Neurosciences 24, 517-526). Slowing ofactivation is likely to delay the onset of action potentialrepolarisation; slowing of deactivation could lead to hyperpolarisingcurrents that reduce the excitability of the neuron and delay the timebefore the neuron can fire a further action potential. Together theseslowing effects on channel activation and deactivation are likely tolead to a reduction rather than a facilitation of the neurons ability tofire at high frequencies. Thus compounds that have this slowing effecton the Kv3.1 and/or Kv3.2 channels may slow neuronal firing. Thisslowing of neuronal firing by a compound of the invention, specificallyExample 15 which markedly increases Tau_(act) to 50.1±7.5 msec (Table1), can be observed from recordings made from “fast-firing” interneuronsin the cortex of rat brain, using electrophysiological techniques, invitro. As can be observed in FIG. 2, the addition of Example 15 reducesthe ability of the neurons to fire in response to trains of depolarisingpulses at 300 Hz.

FIG. 2 shows recordings made from identified “fast-firing” interneuronsin the somatosensory cortex of the mouse. The neurons are induced tofire at high frequencies by trains of high frequency depolarisingcurrent pulses at 100, 200, and 300 Hz. The ability of the neuron tofire an action potential on each pulse is determined. A spikeprobability of 1 on the y-axis of the graph indicates that an actionpotential is generated by the neuron on each of the depolarising currentpulses. In the absence of drug (closed circles, n=9), the neuronsmaintained a spike probability of 1 up to 300 Hz. However, in thepresence of Example 15 (1 microM; open circles, n=6), the neurons wereunable to follow trains at the highest frequency. * p<0.05, ANOVA forrepeated measures.

Therefore, although all the Examples herein identified act as positivemodulators in the recombinant cell assay of Biological Example 1, thosecompounds which markedly increase the value of Tau_(act), such asExample 15, may reduce the ability of neurons in native tissues to fireat high frequency.

In one aspect of the invention, there is provided a Kv3 potentiatingcompound which is associated with a mean tau value that is not more that2 standard deviations greater than the mean value obtained in thepresence of vehicle (DMSO 0.5%), for use in the treatment of disorderswhere positive modulation of Kv3.1 and/or Kv3.2 channel function isbeneficial, including schizophrenia, bipolar disorder, hearingdisorders, sleep disorders, substance-related disorders, and epilepsy.

In one aspect of the invention, there is provided a Kv3 potentiatingcompound which is associated with a mean tau value that is more that 2standard deviations greater than the mean value obtained in the presenceof vehicle (DMSO 0.5%), for use in the treatment of disorders whereinhibition of Kv3.1 and/or Kv3.2 channel function is beneficial,including hyperacusis, Fragile-X, and autism.

Preclinical Experiments

All in vivo studies were conducted in compliance with Project Licensesobtained according to Italian law (art. 7, Legislative Decree no. 116,27 Jan. 1992), which acknowledged the European Directive 86/609/EEC, andwith the GlaxoSmithKline company policy on the care and use oflaboratory animals and related codes of practice.

In the studies that follow, Compound 48 is the compound of ReferenceExample RE1.

Biological Example 2 Evaluation of Compound Effects on the Firing ofInterneurons in the Somatosensory Cortex of Mice, In Vitro Animals

Transgenic mice [CB6-Tg (Gad1-EGFP) G42Zjh/J] were purchased from TheJackson Laboratory (Maine, USA). These mice selectively express enhancedgreen fluorescent protein (EGFP) in the calcium-binding proteinparvalbumin (Pv)-expressing subclass of basket interneurons. EGFPexpression is not reported in other interneuron classes positive forsomatostatin (SOM), cholecystokinin (CCK), calretinin (CR), and VIP.These mice are therefore useful for the identification of thePv-expressing subset of GABAergic neurons that express Kv3.1 and Kv3.2channels and are able to fire at high frequency.

Slice Preparation

Experiments were performed on 250-μm-thick brain slices containing thesomatosensory cortex. Briefly, brains were removed from deeplyanaesthetized (isofluorane) 25-35 day-old Gad1-EGFP mice. Slices werecut using a DTK 1000 microslicer (DSK, Japan) in the following solution(in mM): KCl (2.5), CaCl₂ (0.1), NaH₂PO₄ (1.2), MgCl₂ (5), NaHCO₃ (26),sucrose (189) and glucose (10), kept at 2-6° C. and gassed with 95%O₂-5% CO₂. After cutting, the slices were left to equilibrate in arecovery chamber for at least one hour in an artificial cerebrospinalfluid (ACSF) containing (in mM): NaCl (120), KCl (2.5), CaCl₂ (2),NaH₂PO₄ (2.5), MgCl₂ (1.5), NaHCO₃ (26), and glucose (10), at roomtemperature and saturated with 95% O₂-5% CO₂.

Electrophysiological Recordings

For electrophysiological recordings, a slice was transferred to asubmersion chamber mounted on the stage of an upright microscope(Axioskop, Carl Zeiss, Germany) and superfused with oxygenated ACSF.Visualization of neurons in the slices was accomplished with a 40×objective using infrared-differential interference contrast (IR-DIC)video microscopy (Hamamatsu C5985, Hamamatsu City, Japan).Parvalbumin-positive interneurons were identified by illuminating thepreparation with a fluorescence lamp with a GFP-filter and switchingbetween fluorescence and IR-DIC video microscopy. Only GFP-positiveneurons were recorded. Whole-cell recordings were made usingborosilicate-glass patch pipettes pulled using a Sutter P-97 electrodepuller and filled with an internal solution containing (in mM):KGluconate (125), EGTA (10), HEPES (10), MgCl₂ (1), KCl (10) and MgATP(2); pH 7.3 adjusted with KOH. When filled with this internal solution,patch electrodes had a tip resistance of 4-7 M. Recordings were carriedout at room temperature (20-22° C.) using a Multiclamp 700B amplifier(Axon Instruments, Foster City, Calif., USA). Current-command protocols(indicated below) and data acquisition were performed using pClamp 10.0software and a Digidata 1320A interface (Axon Instruments, Foster City,Calif., USA). Capacitive transients were neutralised andseries-resistance was monitored continuously throughout the experiment.If it changed by >20% the cell was discarded. Data were filtered at 3kHz and sampled at 10 kHz.

Drugs

Compounds of the invention were dissolved in DMSO (100%),tetraethylammonium (TEA) and tetrodotoxin (TTX), (both from Sigma,Italy) were dissolved in distilled water and stored at −20° C. untiluse. Drugs were diluted to the final concentration on the day of theexperiment. The highest final concentration of DMSO used was 0.1%.

Experimental Procedure

The firing activity of the recorded interneurons was evaluated byapplying long current steps at different intensities. Thus, after theformation of a giga-seal, the amplifier was switched to current-clampmode, allowing the neuron to reach its resting membrane potential. Anegative current was then injected into the cell in order to obtain aresting potential close to −80 mV. From this condition, step currentinjections (50 pA increments, 600 ms) were applied to elicit actionpotentials. This protocol was repeated at least 2 times for each cell.

Online bridge-balance compensation was carried out and R_(m) value wasmonitored continuously throughout the experiment.

Drug Application

Slices were incubated in the recovery chamber for at least 1 hour in thepresence of either vehicle (0.1% DMSO), TEA (0.5 mM)+0.1% DMSO, or TEA(0.5 mM)+Reference Example RE1 (1 or 10 microM). After transfer of aslice to the recording chamber, the same drug condition was maintainedby superfusion of the appropriate drugs in the circulating ACSF.

Data Acquisition and Analysis

Raw data were acquired using Clampex 10.0 (Molecular Devices, USA). Datawere analyzed using Clampfit 10.0 software (Molecular Devices, USA). Thefrequency of action potential firing (expressed in Hz) in response tostep current injections was calculated from the number of actionpotentials detected over the 600 ms step current. Values of frequencyobtained for each current step in the same experimental condition and inthe same cell were averaged. Since the threshold to evoke actionpotentials differed from one cell to another, current step intensity wasexpressed as pA from the current threshold for action potentialgeneration, rather than in absolute values.

Action potential half-width was calculated for each action potentialusing Clampfit. The values of the 2^(nd)-5^(th) or the last ten actionpotentials evoked by a non-saturating current step (typically 100-150 pAfrom threshold) were averaged for each experimental condition in eachanalyzed cell.

Statistical Analysis

Statistical differences between the effect of treatments on actionfiring frequency were evaluated using a two-way ANOVA for repetitivemeasurements and, if necessary, post hoc planned comparisons(differences were considered significant where p<0.05). The effect ofdrug treatment on action potential half-width and on the firstderivative amplitude was evaluated using an ANOVA. All statisticalanalyses were conducted using Statistica Software (StatSoft version 8).When appropriate, results were reported as mean±SEM.

Criteria for Data Inclusion/Exclusion

The criteria used to include or exclude a cell from the analysis werebased on accurate current-clamp conditions and the stability of therecording throughout the experiment. Online evaluation allowed theexclusion of a cell when the R_(s) and/or R_(m) values changed by >20%.

Results

Interneurons recorded from slices incubated with 0.5 mM TEA fired at alower maximal frequency in response to step currents compared to neuronsrecorded from control slices (FIG. 3). This effect was significantlyreversed in slices incubated with TEA (0.5 mM) plus Reference ExampleRE1 at 1 μM or 10 μM (one-way ANOVA for repeated measurements, * p<0.05with respect to TEA alone).

FIG. 3. The frequency of action potentials recorded fromparvalbumin-positive interneurons in the somatosensory cortex of themouse, evoked by depolarizing current steps (600 ms duration andΔ-increment of 50 pA) after at least 1 hour with either vehicle (0.1%DMSO; filled circles, n=6), TEA (0.5 mM)+0.1% DMSO (open circles, n=7),TEA (0.5 mM)+Reference Example RE1 (1 μM; filled triangles, n=9), or TEA(0.5 mM)+Reference Example RE1 (10 μM; open triangles, n=5). * p<0.05;One-way ANOVA for repeated measurements.

Furthermore, the action potential half-width and was significantlyincreased in cells recorded from slices incubated with TEA (0.5 mM)compared to control slices (0.1% DMSO) (FIG. 4). In slices incubatedwith TEA (0.5 mM) plus Reference Example RE1 at 1 μM or 10 μM, the meanaction potential half-width was significantly decreased by 24% and 36%,respectively, compared to slices incubated with TEA (0.5 mM) only (ANOVAand Dunnett test, * p<0.05, n=9; ** p<0.01, n=5, respectively).

FIG. 4. The half-width of evoked action potentials fromparvalbumin-positive interneurons in the somatosensory cortex of themouse. Prior to recordings, slices were incubated for at least 1 hourwith either vehicle (Control; 0.1% DMSO, n=6), TEA (0.5 mM)+0.1% DMSO(n=7), TEA (0.5 mM)+Reference Example RE1 (1 μM; n=9), or TEA (0.5mM)+Reference Example RE1 (10 μM; n=5). * p<0.05; ** p<0.01, ***p<0.001, ANOVA followed by Dunnett test.

These results demonstrate the ability of compounds which have activityin the assays of Biological Example 1 to modulate the behaviour offast-firing interneurons in the mouse brain in a manner consistent withpositive modulation of Kv3.1 and/or Kv3.2 channels. The ability toenhance Kv3 function in cortical brain areas is also consistent with thepotential of these compounds to treat a range of central nervous system,disorders, including schizophrenia, bipolar disorder, and epilepsy.

Biological Example 3 Evaluation of Compound Effects on PotassiumCurrents Recorded from Neurons in the Medial Nucleus of the TrapezoidBody in Mice, In Vitro Animals

Male CBA/Ca mice (aged 12-16 days) were used in these experiments (inaccordance with the UK Animals Scientific Procedures Act, 1986). Brainslices containing the medial nucleus of the trapezoid body (MNTB) wereprepared as described previously (Brew and Forsythe, 2005).

Drugs

Chemicals and reagents were purchased from Sigma, (Poole, UK) unlessotherwise noted. Reference Example RE1 was dissolved in DMSO and dilutedin ACSF to the required concentration.

Electrophysiological Recording

Recordings from identified MNTB neurons were conducted as previouslydescribed (Brew and Forsythe, 2005). Slices was placed in a superfusionchamber on an inverted microscope stage and continuously perfused withgassed (95% O₂-5% CO₂) ACSF at a rate of 1 ml min⁻¹ at room temperature.Whole-cell recordings were made from visually identified MNTB neuronsusing an Axopatch 700B amplifier (Molecular Devices, Union City, Calif.,USA). Patch solution comprised (in mm) potassium gluconate (97.5), KCl(32.5), Hepes (40), EGTA (5), MgCl₂ (1), Na₂phosphocreatin (5), pH 7.2with KOH. Pipettes had resistances of 3-5 MΩ and series resistances were6-10 MΩ (compensated by 70%, 10 μs lag). Access resistance wasfrequently monitored and the recording discarded if increases were morethan 2 MΩ.

Once a whole-cell configuration had been obtained, cells were held at−60 mV prior to application of voltage protocols as follows: cells werestepped from the holding potential to −90 for 700 ms and stepped to −40mV for 25 ms and then a voltage pulse to a range of voltages from −100to +40 mV (10 mV increments) was applied for 220 ms before returning tothe holding potential. Following completion of this protocol, TEA (1 mM)was added to the superfusion medium. After 5 minutes, a second set ofrecordings using the same voltage protocol was carried out. Followingthis, Reference Example RE1 (10 microM) was added to the ACSF, in thecontinuing presence of TEA (1 mM), and after a further 5 minutes, afinal set of recordings with the voltage protocol was made.

Statistical Analysis

Currents evoked by the voltage step to +40 mV were compared across drugtreatments for each cell using an unpaired t-test.

Results

TEA (1 mM) significantly reduced the amplitude of outward, highvoltage-activated potassium currents evoked by voltage steps to +40 mV(FIG. 5). This effect was reversed by the subsequent application ofReference Example RE1 (10 microM).

FIG. 5. High-voltage activated potassium currents recorded from visuallyidentified MNTB neurons in the mouse, in vitro. Data shown are the mean(+/−s.d.) of the current amplitude evoked by voltage steps to +40 mVunder different drug conditions. TEA (1 mM), TEA (1 mM)+ReferenceExample RE1 (10 microM). Statistical analysis was conducted using anunpaired t-test.

These data indicate that compounds which have activity in the assays ofBiological Example 1 can modulate high voltage-activated potassiumcurrents (presumed to be mediated by Kv3.1 channels; Brew and Forsythe,2005) in neurons of the MNTB, a region of the brainstem that processesauditory information. This result supports the utility of compounds ofthe invention for the treatment of hearing disorders.

Biological Example 4 Electroshock Seizure Model in Rats ExperimentalPreparation

Male CD rats (85-130 g) were supplied by Charles River, Italy. Animalswere group housed with free access to food (Standard rodent chow) andwater under a 12 h light/dark cycle (lights on at 0600 h). A period ofat least 5 days between arrival at GSK and the study was allowed in allcases.

Experimental Protocol

Animals were administered a test compound at the appropriate dose, routeand pre-treatment time and returned to their home cage. Testing occurredin a separate room from that used for housing. Testing involveddetermining the threshold for tonic hindlimb extensor seizures using aHugo Sachs Electronik stimulator which delivers a constant current of0.3 second duration, 50 Hz, sinewave form, fully adjustable between 1and 300 mA. Stimuli were delivered via corneal electrodes (Stean T O,Atkins A R, Heidbreder C A, Quinn L P, Trail B K, Upton N. (2005) Br JPharmacol. 144(5):628-35). Seizure threshold was determined using the‘up and down’ method of Kimball et al. (1957) (Kimball A W, Burnett W TJr, Doherty D G. (1957) Radiat Res. 7(1):1-12). The first animal testedin each group was stimulated with a current that might be expected to beclose to the threshold for induction of a seizure. If a tonic seizurewas not induced, then the next animal in the group received a stimulus 5mA higher. If a tonic seizure was induced, then the next animal receiveda stimulus 5 mA lower. This is repeated for all animals within thecontrol (vehicle) group. In the case of groups treated with a testcompound steps of 5 to 10 mA were used. At the end of the study, bloodsamples were taken for analysis of the drug concentrations in thiscompartment (n=4/group).

Drugs and Materials

All doses were calculated as base. Sodium valproate was suspended inMethocell 1% (w/v) and dosed via the oral (p.o.) route at 5 mL/kg 1 hourbefore test. Reference Example RE1 was dissolved in DMSO and thensuspended in Methocell 1% (w/v) to a final DMSO concentration of 5%(v/v). Reference Example RE1 was then dosed p.o. at 5 mL/kg 2 hoursbefore test.

Data Analysis

Induction of seizure is measured as an all-or-nothing effect scored aseither present (+) or absent (0) for each animal. The data for eachtreatment group were recorded as the number of +'s and 0's at eachcurrent level employed and this information was then used to calculatethe CC50 value (current required for 50% of animals to show seizurebehaviour)+standard error of the mean according to the method of Kimballet al. (1957). Drug effects were calculated as the % change in CC50.Significant differences between drug-treated animals and appropriatevehicle treated groups were assessed according to the methods ofLitchfield and Wilcoxon (1949).

Results

Pretreatment with Reference Example RE1 was associated with asignificant increase in seizure threshold at both doses tested: At thedose of 30 mg/kg p.o., Reference Example RE1 produced a 91% increase inseizure threshold, whereas at the dose of 60 mg/kg p.o., the increase inseizure threshold was +218%. The increase produced by the higher dose ofReference Example RE1 was similar to the increase produced by thepositive control, sodium valproate at 300 mg/kg p.o. (+258%).

Blood concentrations of Reference Example RE1 measured in satelliteanimals 2 hours after dosing were 5.3 and 9.1 μg/mL following the dosesof 30 and 60 mg/kg p.o., respectively. These concentrations areequivalent to unbound concentrations in blood of 1.3 and 2.2 μM,respectively, and thus are consistent with concentrations of ReferenceExample RE1 that produce a significant increase in Kv3-mediated currentsobserved in the in vitro recombinant human Kv3 electrophysiology assay,described above.

Conclusions

These results suggest that Reference Example RE1 has anticonvulsantefficacy, and that this effect is likely to be mediated by the positivemodulation of Kv3 potassium channels. Consequently, compounds which haveactivity in the assays of Biological Example 1 can have anticonvulsantefficacy.

Biological Example 5 Psychostimulant-Induced Hyperactivity in MiceExperimental Preparation

Male CD-1 mice (25-35 g) were supplied by Charles River, Italy. Animalswere group housed with free access to food (Standard rodent chow) andwater under a 12 h light/dark cycle (lights on at 0600 h). A period ofat least 5 days between arrival at GSK and the study was allowed in allcases.

Experimental Protocol

Animals were administered a test compound at the appropriate dose, routeand pre-treatment time, and then returned to their home cage. Testingoccurred in a separate room from that used for housing. Mice weretreated orally (p.o.) with the test compound and placed individuallyinto a Perspex box (length 20.5 cm, width 20.5 cm, height 34 cm) coveredwith a perforated lid. Infrared monitoring sensors were located aroundthe perimeter walls (horizontal sensors). Two additional sensors werelocated 2.5 cm above the floor on opposite sides (vertical sensors).Data were collected and analysed using a VersaMax System (AccuscanInstruments Inc., Columbus, Ohio) which in turn transferred informationto a computer. After 30 minutes of habituation, mice were treated withamphetamine dosed intraperitoneally (i.p.) at 2 mg/kg at 10 mL/kg, andsubsequent locomotor activity in the test arena was assessed over afurther 60 minutes. Locomotor activity was determined as the totaldistance (cm) traveled by each mouse in the test arena over the 60minute test period.

Drugs and Materials

All doses were calculated as base. Clozapine was dissolved in distilledwater and dosed at 3 mg/kg intraperitoneum (i.p.) at 10 mL/kg. ReferenceExample RE1 (10, 30 or 60 mg/kg) or vehicle (HPMC 0.5% w/v, Tween80 0.1%v/v in water) was administered p.o. at 10 mL/kg. Both clozapine andReference Example RE1 were dosed immediately before placing the animalin the test arena (30 minutes before amphetamine administration).

Results

Amphetamine alone produced a large and significant increase in totaldistance traveled. A dose of 30 mg/kg p.o. of Reference Example RE1significantly reduced the increase in total distance traveled producedby amphetamine. A higher dose of 60 mg/kg p.o. of Reference Example RE1further reduced the increase in locomotor activity induced byamphetamine in a manner similar to the positive control, clozapine (3mg/kg i.p.). Data are summarised in Table 1.

TABLE 1 Effects of Reference Example RE1 on amphetamine inducedhyperlocomotion in the mouse. Reference Example RE1 was administeredp.o. 30 minutes before amphetamine (2 mg/kg i.p.). Clozapine wasadministered i.p. 30 minutes before amphetamine (2 mg/kg i.p.). Totaldistance was assessed over 60 minutes starting immediately afteramphetamine administration. Data are expressed as mean ± sem. Data weresubjected to one-way analysis of variance (ANOVA) followed by Dunnett'stest (** = p < 0.01 vs amphetamine treatment alone). Treatment TotalDistance Travelled (cm) Vehicle 1049 ± 522** Amphetamine (AMPH) 2.0mg/kg 16304 ± 3309 AMPH 2 mg/kg + Reference Example RE1 10 mg/kg 15267 ±3166 AMPH 2 mg/kg + Reference Example RE1 30 mg/kg 5790 ± 1436** AMPH 2mg/kg + Reference Example RE1 60 mg/kg 1494 ± 378** AMPH 2 mg/kg +Clozapine 3 mg/kg 932 ± 362**

Conclusions

These results show that Reference Example RE1, at doses similar to thosethat show anticonvulsant efficacy, is able to prevent hyperactivityinduced by the psychostimulant, amphetamine. Thus, reference Example RE1and other compounds that positively modulate Kv3.1 and/or Kv3.2channels, as can be observed from the assay described in BiologicalExample 1, may be useful in the treatment of disorders associated withhyperactivity, such as bipolar mania, or disruption of the dopaminesystem, such that may occur in drug dependence, attention deficithyperactivity disorder (ADHD), or schizophrenia.

Biological Example 6 Pharmaco-Electroencephalography (phEEG) in theCommon Marmoset Animals and Surgery

Laboratory bred male (vasectomised) and female common marmosets(Callithrix jacchus) over 2 years of age, weighing 250-500 g were usedin this study. The animals were caged in couples, in a housing roommaintained at 25±1° C., 60% humidity and a 12 hour light/dark cycle(lights on at 0600, with 30 min simulated dawn and twilight). Animalsreceived a standard diet and drinking water ad libitum. Only one animalof each pair was involved in the test, which was carried out with theanimal situated in the home cage.

The effect of compounds of the invention was assessed using telemetricrecording of cortical EEG (ECoG). A multichannel telemetric transmitter(DSI model TL11M2-F40-EET) is implanted intraperitoneally using standardsurgical techniques in anaesthetised marmosets. Recording electrodeswere permanently fixed, with dental cement, to the skull directly incontact with the dura mater through two drilled holes in thefronto-parietal region. Following surgery, animals were housed in pairs(one implanted, one unoperated partner) in their home cage with accessto food and water ad libitum. Animals demonstrated a normal behaviouralrepertoire immediately after recovery from surgery; however, phEEG wasassessed at least 3 weeks later. All in vivo studies were conducted inaccordance with the Italian laws and conformed to GlaxoSmithklineethical standards.

Experimental Procedure

The animals were placed in the nest-boxes in their room cages and EEGtraces were recorded using Dataquest ART software for a 5-min period foreach time-point and analyzed using Spike2 software (CED, UK). Thespectral power in each frequency band was determined for each 2 secepoch during the pre-treatment period and averaged; similarly spectralpower in each band was determined for successive 2 sec epochs of each5-min period of recording following vehicle or drug treatment. Change inthe absolute spectral power, for each of the different bands (delta,theta, alpha and beta) was calculated offline.

Drug treatments were assigned according to a complete crossover design:All treatments were randomly distributed between animals, in separateexperimental sessions, each animal received vehicle and each dose ofdrug, after an appropriate wash-out period.

Six animals were treated orally with Reference Example RE2 at the dosesof 0.3, 1 and 3 mg/kg (1 ml/kg) and the EEG traces were recorded at +15,30, 60, 90, 120 and 180 minutes following treatment. Reference ExampleRE2 was suspended in 12.5% (w/v) aqueous captisol containing 0.1% (w/v)Tween80 and 0.5% (w/v) HPMC.

Data Analysis

Four different frequency bands were considered: delta (1.50-6.00 Hz),theta (6.00-8.00 Hz), alpha (8.00-12.00 Hz) and beta (12.00-30.00 Hz).Values for spectral power in each band at each time point were first logtransformed and then analysed with a mixed effect model with time asfixed effect, the baseline level as covariate, and animal as randomterm. Data are summarised as mean of the percentage changes frombaseline and standard error.

Results

The pharmaco-EEG changes observed in these studies show that, comparedto vehicle, Reference Example RE2 at the highest dose (3 mg/kg) induceda statistically significant increase of the absolute power in the deltaband between 30 and 120 minutes (p<0.05) and a statistically significantincrease in theta band power at 60 minutes (p<0.05). At the intermediatedose (1 mg/kg) Reference Example RE2 induced a marginally significant(p<0.10) increase in the absolute power in delta band at 30 minutes anda concomitant significant reduction in the beta band (p<0.05). Nosignificant effects were observed in the alpha band at any dose ofReference Example RE2.

These results suggest that compounds which have activity in the assaysof Biological Example 1 can modify the EEG of awake primates. Increasesin delta-band EEG activity have previously been observed withantipsychotic compounds in humans.

Biological Example 7 Circadian Pattern of Expression of Kv3.1 and Kv3.2Channels in the Superchiasmatic Nucleus of Mice Material & Methods

Thirty adult male C57BL/6J mice (age: 4-5 weeks at the arrival; CharlesRiver FR) were stored in six different cages (5 mice per cage) andmaintained for 4 weeks in a dedicated room with 12 h light-12 h darkcondition (lights on at 06:00, designated as Circadian time [CT] 6;lights off at 18:00, designated as CT 18. The room temperature wasmaintained at 21±2° C.; food and water were available ad libitum.

After this period, mice were sacrificed by cervical dislocation atdifferent time points over a 24 hour period; 5 mice per time point. Theanimals were transferred from the storage room to the surgery room andthen immediately sacrificed. During the dark phase, all these actionswere performed under a dim red light.

Brains were removed from the skull and immediately immersed inisopentane maintained approximately at −30° C. and then stored at −70°C. prior to in situ hybridization analysis.

All the brains were cut by cryostat and a number of 14 μm-thick coronalsections were collected at the level in which the suprachiasmaticnucleus is present, approximately between −0.22 mm and −0.82 mm fromBregma (Paxinos and Franklin, “The mouse brain in stereotaxiccoordinates”). The sections were then stored at −80° C. until usage. Foreach time point (CT12, 16, 20, 24, 4 and 8), five mice were collectedand two non-consecutive sections for each mouse were selected andallowed to dry at room temperature, then immediately exposed to in situhybridization protocol, as described in the previous experiments.

Results

A one-way ANOVA found no significant effect of time on Kv3.1 mRNAexpression within the mouse suprachiasmatic nucleus (FIG. 6 a). Incontrast, one-way ANOVA indicated that there was a highly significanteffect of the time on Kv3.2 mRNA expression within the mousesuprachiasmatic nucleus (p<0.001), with a significant peak of expressionat ZT 10, the time point corresponding to 2 hours before the shift fromthe light to the dark phase, which is the active phase of the mice (FIG.6 b).

FIG. 6: (a) Expression of Kv3.1 mRNA in the suprachiasmatic nucleus ofmice sacrificed during at different Circadian times over a 24-hourlight-dark cycle. Kv3.b1 mRNA expression is expressed in nCi/g asmean±S.E.M. from n=5 mice per time point. (b) Expression of Kv3.2 mRNAin the superchiasmatic nucleus. Kv3.2 mRNA expression is expressed asmean±S.E.M. from n=5 mice per time point. *** p<0.001: Kv3.2 mRNAexpression at CT 16 is significantly different from all the othertimepoints. * p<0.05: the Kv3.2 mRNA expression at CT 20 issignificantly different from the expression measured at CT 24 and 4.

These results indicate that Kv3.2 channel expression in thesuperchiasmatic nucleus varies over the 24-hour circadian cycle. Thus,given the central role of the superchiasmatic nucleus in setting thecircadian clock in mammals, Kv3.2 channels are likely to be important tothe function of this clock. Consequently, compounds that modulate Kv3.2channels may have potential in the treatment of disorders associatedwith circadian dysfunction, including sleep and bipolar disorders.

Biological Example 9 Assessment of Physiological Sleep in the RatMethods:

Adult male CD rats (C. River, Italy) were implanted with telemetricprobes and housed singly, under controlled condition (temperature 18-20g C; relative humidity 45-50%; 12 hours light-dark cycle, lights on at 3p.m., designated Circadian Time (CT) 0) with free access to food andwater.

Reference Example RE1 was formulated in tween (0.1% v/v) and HPMC (0.5%v/v) and was administered orally at doses of 10, 30 and 60 mg/kg (vol. 2ml/kg) at CT18 (6 hours before lights on). The electroencephalogram(EEG) and electromyogram (EMG) were recorded continuously, startingimmediately after administration, using telemetric apparatus (DSIDataquest® A.R.T. system). EEG and EMG recordings were analysed toevaluate sleep patterns using sleepSign® (Kissei Comtec Co.).Statistical analyses (1-way Anova followed by a Dunnett's test) wasperformed using Statistica-8 software.

Results:

Reference Example RE1 at 60 mg/kg significantly increased total sleeptime (p<0.05, n=8) and time spent in non-REM sleep (p<0.05, n=8) overthe 5 hour period immediately following dosing, but did not affect timespent in REM sleep.

These results suggest that compounds of the invention can increasephysiological sleep in animals, which suggests that they may be usefulin the treatment of sleep disorders in humans.

1.-43. (canceled)
 44. A compound of formula (I):

wherein: R₁ is H, or C₁₋₄alkyl, halo, haloC₁₋₄alkyl, CN, C₁₋₄alkoxy, haloC₁₋₄alkoxy; R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or halo; R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo; or R₃ is absent; R₁₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo; or R₁₃ is absent; A is a 5 or 6 membered saturated or unsaturated heterocycle, with at least one O atom; which heterocycle is optionally fused with a cyclopropyl group to form a tricycle when considered together with the phenyl; X is C or N; Y is C or N; R₄ is C₁₋₄ alkyl; R₅ is H, Deuterium, C₁₋₄ alkyl; or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly; wherein R₂ and R₃ may be attached to the same or a different ring atom; and wherein R₂ may be attached to a fused ring atom; or a pharmaceutically acceptable salt and/or solvate thereof.
 45. The compound according to claim 44, which is:

wherein R₁ is H, or C₁₋₄alkyl, halo, haloC₁₋₄alkyl, CN, C₁₋₄alkoxy, haloC₁₋₄alkoxy; R₂ is H, C₁₋₄alkyl, C₃₋₄ spiro carbocycly, haloC₁₋₄alkyl or halo; R₃ is H, C₁₋₄alkyl, haloC₁₋₄alkyl, halo; A is a 5 or 6 membered saturated or unsaturated heterocycle, with at least one O atom; which heterocycle is optionally fused with a cyclopropyl group to form a tricycle when considered together with the phenyl; X is C or N; Y is C or N; R₄ is C₁₋₄ alkyl; R₅ is H, Deuterium, C₁₋₄ alkyl; or R₄ and R₅ can be fused to form C₃₋₄ spiro carbocycly; wherein R₂ and R₃ may be attached to the same or a different ring atom; and wherein R₂ may be attached to a fused ring atom; or a pharmaceutically acceptable salt thereof.
 46. The compound according to claim 44, wherein R₁ is H or methyl.
 47. The compound according to claim 44, wherein A is a 5 membered saturated or unsaturated heterocycle, with at least one O atom; which heterocycle is optionally fused with a cyclopropyl group to form a tricycle when considered together with the phenyl.
 48. The compound according to claim 44, wherein A is a 6 membered saturated or unsaturated heterocycle, with at least one O atom; which heterocycle is optionally fused with a cyclopropyl group to form a tricycle when considered together with the phenyl.
 49. The compound according to claim 44, wherein the ring A is dihydrofuran or dihydropyran.
 50. The compound according to claim 44, wherein R₂ is H, methyl or a C₃ spiro group.
 51. The compound according to claim 44, wherein R₃ is H, F, methyl or ethyl.
 52. The compound according to claim 51, wherein R₃ is H or methyl.
 53. The compound according to claim 44, wherein R₁₃ is H, F or methyl.
 54. The compound according to claim 44, wherein R₁₃ is absent.
 55. The compound according to claim 44, wherein X is C and Y is C.
 56. The compound according to claim 44, wherein X is C and Y is N.
 57. The compound according to claim 44, wherein X is N and Y is N.
 58. The compound according to claim 44, wherein R₄ is methyl or ethyl.
 59. The compound according to claim 44, wherein R₅ is H or methyl.
 60. The compound according to claim 44, wherein R₄ and R₅ have the stereochemical arrangement:


61. A method for the prophylaxis or treatment of hearing disorders, schizophrenia, bipolar disorder, epilepsy or sleep disorders by administering to a subject a compound according to claim
 44. 62. A compound according to claim 44, in the form of a pharmaceutical composition comprising said compound and a pharmaceutically acceptable carrier or excipient.
 63. A compound selected from: spiro[1-benzofuran-3,1′-cyclopropan]-4-ol

7-methylspiro[2H-benzofuran-3,1′-cyclopropane]-4-ol

and 3,3,7-trimethyl-2,3-dihydro-1-benzofuran-4-ol

or a salt thereof; or a compound selected from:

wherein: X is C or N; Y is C or N; and the group W is selected from:

or a salt thereof. 